Biguanide derivatives and their rearrangement products for use in the treatment of cancer

ABSTRACT

The present disclosure relates to biguanide derivatives of formula (I) and their rearrangement products. The present disclosure also relates to the use of these compounds in a method for treating cancer, in particular melanoma.

TECHNICAL FIELD

The present disclosure relates to biguanide derivatives and theirrearrangement products. The present disclosure also relates to the useof these compounds in a method for treating cancer, in particularmelanoma.

BACKGROUND

Cancers represent one of the most important causes of death in France.Among cancers, melanoma is a skin cancer that is widespread in France,with about 8000 new cases diagnosed each year and more than 1000 deaths.This cancer is therefore a major public health problem. Melanoma is amalignant tumor developed from melanocytes that are responsible for thesynthesis of melanin, which are photoprotective pigments. Melanoma is anextremely aggressive tumor with a high metastatic potential towardslymph nodes, liver, lungs, central nervous system and skin. As soon asmetastases appear, the vital prognosis becomes unfavorable because ofthe poor efficiency of all current treatments.

Recently, encouraging results have been obtained with BRAF inhibitors(vemurafenib (PLX 4032) and dabrafenib), which target only B-Raf mutantmelanomas (approximately 50% of the metastatic melanoma). Unfortunately,after a short period of remission, melanoma, in almost all cases, gainsresistance against these drugs and metastases develop again, increasingthe patient's life expectancy by about 2 months. Immunotherapies havealso been developed recently. They are based on anti-CTLA4 and Anti-PD1antibodies that reactivate the immune response. However, immunotherapiesgive an objective response in only 15 to 30% of patients.

It has also been reported that metformin, a drug normally used to treatdiabetes, is able to inhibit the growth of melanoma cells (Tomic et al.,Cell Death and Disease, 1; 2: e199, 2011) and the development ofmetastasis (Cerezo et al, Molecular Cancer Therapeutics, 12 (8):1605-15, 2013). Nevertheless, a high dose of metformin is likely to berequired to induce cell death (IC50=10 mM, ten millimolar).

Thus, there is a need for compounds and compositions withanti-proliferative properties, which could be used in particular totreat patients with melanoma and for example, patients with BRAFinhibitor-resistant melanoma.

SUMMARY

The inventors surprisingly found that biguanide derivatives comprising aheteroaryl moiety, and their rearrangement products, have a highbiological activity towards cancer lines such melanoma cell lines,including melanoma cell lines resistant to BRAF inhibitors.

Consequently, in a first aspect, the disclosure relates to a compound offormula (I)

-   -   wherein    -   R₁ and R₂ are independently selected from H, halogen, C₁-C₆        alkyl, C₃-C₆ cycloalkyl, C₁-C₆ haloalkyl, C₂-C₆ alkenyl, C₂-C₆        alkynyl, heterocyclyl having 5 to 10 ring atoms, aryl having 6        to 10 ring atoms, heteroaryl having 5 to 10 ring atoms, and        C₇-C₁₆ aralkyl, said alkyl, cycloalkyl, haloalkyl, alkenyl,        alkynyl, heterocyclyl, aryl, heteroaryl and aralkyl being        optionally substituted with one or more substituents        independently selected from oxo, halogen, C₁-C₆ alkyl, C₁-C₆        alkoxy, C₁-C₆ haloalkyl, —OH, —NR″R′″, —NO₂, —CN and —(CO)—R;    -   or R₁ and R₂, together with the carbon-carbon double bond        between them, form a 6 to 10 membered aryl or heteroaryl ring,        said aryl and heteroaryl being optionally substituted with one        or more substituents independently selected from oxo, halogen,        C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —OH, —NR″R′″, —NO₂,        —CN and —(CO)—R;    -   Y is −O— or —S—;    -   R₃ is selected from C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₁-C₆        haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, heterocyclyl having 5        to 10 ring atoms, aryl having 6 to 10 ring atoms, heteroaryl        having 5 to 10 ring atoms and C₇-C₁₆ aralkyl, said alkyl,        cycloalkyl, haloalkyl, alkenyl, alkynyl, heterocyclyl, aryl,        heteroaryl and aralkyl being optionally substituted with one or        more substituents independently selected from oxo, halogen,        C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —OH, —NR″R′″, —NO₂,        —CN and —(CO)—R;    -   each R is independently selected from H, C₁-C₆ alkyl, C₁-C₆        alkoxy and —NR″R′″;    -   each R″ and R′″ is independently selected from H and C₁-C₆        alkyl.

In a second aspect, the disclosure relates to a compound of formula (II)or (III)

-   -   wherein    -   Y′ is —SR₄ or —OR₅,    -   R₄ is selected from H, C₁-C₆ alkyl and protecting groups;    -   R₅ is selected from H and protecting groups;    -   each R′ is independently selected from halogen, C₁-C₆ alkyl,        C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —OH, —NR″R′″, —NO₂, —CN and        —(CO)—R;    -   n is 0 to 4;    -   R₃ is selected from C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₁-C₆        haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, heterocyclyl having 5        to 10 ring atoms, aryl having 6 to 10 ring atoms, heteroaryl        having 5 to 10 ring atoms, and C₇-C₁₆ aralkyl, said alkyl,        cycloalkyl, haloalkyl, alkenyl, alkynyl, heterocyclyl, aryl,        heteroaryl and aralkyl being optionally substituted with one or        more substituents independently selected from oxo, halogen,        C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —OH, —NR″R′″, —NO₂,        —CN and —(CO)—R;    -   each R is independently selected from H, C₁-C₆ alkyl, C₁-C₆        alkoxy and —NR″R′″;    -   each R″ and R′″ is independently selected from H and C₁-C₆        alkyl.

In a third aspect, the disclosure relates to a pharmaceuticalcomposition comprising a compound of formula (I), (II) or (III), and apharmaceutically acceptable carrier.

In a fourth aspect, the disclosure relates to a compound of formula (I),(II) or (III) for use in a method for treating cancer.

In a fifth aspect, the disclosure relates to a method for treatingcancer, said method comprising administering to a subject atherapeutically efficient amount of

-   -   (i) a compound of formula (I),    -   (ii) a compound of formula (II) or (III), or    -   (iii) a pharmaceutical composition as described herein.

In a sixth aspect, the disclosure relates to the use of a compound offormula (I), (II) or (III), for the manufacture of a medicament for thetreatment of cancer.

In a seventh aspect, the disclosure relates to the use of a compound offormula (I), (II) or (III) for use as a drug.

DETAILED DESCRIPTION Definitions

As used herein, the terms “C₁-C₆ alkyl”, by itself or as part of anothersubstituent, refer to a linear or branched alkyl functional group having1 to 6 carbon atoms. Suitable alkyl groups include methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl and t-butyl, pentyl andits isomers (e.g. n-pentyl, iso-pentyl), and hexyl and its isomers (e.g.n-hexyl, iso-hexyl).

As used herein, the terms “C₃-C₆ cycloalkyl” refer to a saturated orunsaturated cyclic group having 3 to 6 carbon atoms. Suitable cycloalkylgroups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

As used herein, the term “halogen” refers to a fluoro (—F), chloro(—Cl), bromo (—Br), or iodo (—I) group.

As used herein, the terms “C₁-C₆ haloalkyl” refer to a C₁-C₆ alkyl asdefined herein that is substituted by one or more halogen group asdefined herein. Suitable C₁-C₆ haloalkyl groups include trifluoromethyland dichloromethyl.

As used herein, the terms “C₂-C₆ alkenyl” refer to a straight orbranched hydrocarbon moiety having at least one carbon-carbon doublebond. Alkenyl groups include, for example, ethenyl (i.e., vinyl),propenyl, butenyl, 1-methyl-2-buten-1-yl, pentenyl, hexenyl, octenyl,and butadienyl.

As used herein, the terms “C₂-C₆ alkynyl” refer to a straight orbranched hydrocarbon moiety having at least one carbon-carbon triplebond. Examples of “alkynyl” include ethynyl, 2-propynyl (propargyl),1-propynyl, pentynyl, hexynyl, and allenyl groups, and the like.

As used herein, the terms “C₁-C₆ alkoxy” refer to a —O-alkyl group,wherein the alkyl group is a C₁-C₆ alkyl as defined herein. SuitableC₁-C₆ alkoxy groups include methoxy, ethoxy, propoxy.

As used herein, the terms “aryl having 6 to 10 ring atoms” refer to apolyunsaturated, aromatic hydrocarbyl group having a single ring ormultiple aromatic rings fused together, containing 6 to 10 ring atoms,wherein at least one ring is aromatic. The aromatic ring may optionallyinclude one to two additional rings (cycloalkyl, heterocyclyl orheteroaryl as defined herein) fused thereto. Suitable aryl groupsinclude phenyl, naphtyl and phenyl ring fused to a heterocyclyl, likebenzopyranyl, benzodioxolyl, benzodioxanyl and the like.

As used herein, the terms “heteroaryl having 5 to 10 ring atoms” referto a polyunsaturated, aromatic ring system having a single ring ormultiple aromatic rings fused together or linked covalently, containing5 to 10 atoms, wherein at least one ring is aromatic and at least onering atom is a heteroatom selected from N, O and S. The nitrogen andsulfur heteroatoms may optionally be oxidized and the nitrogenheteroatoms may optionally be quaternized. Such rings may be fused to anaryl, cycloalkyl or heterocyclyl ring. Non-limiting examples of suchheteroaryl, include: furanyl, thiophenyl, pyrrolyl, pyrazolyl,imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl,oxadiazolyl, thiadiazolyl, tetrazolyl, oxatriazolyl, thiatriazolyl,pyridinyl, pyrimidyl, pyrazinyl, pyridazinyl, oxazinyl, dioxinyl,thiazinyl, triazinyl, indolyl, isoindolyl, benzofuranyl,isobenzofuranyl, benzothiophenyl, isobenzothiophenyl, indazolyl,benzimidazolyl, benzoxazolyl, purinyl, benzothiadiazolyl, quinolinyl,isoquinolinyl, cinnolinyl, quinazolinyl and quinoxalinyl.

As used herein, the terms “heterocyclyl having 5 to 10 ring atoms” referto a saturated or unsaturated cyclic group having 5 to 10 ring atoms,wherein at least one ring atom is a heteroatom selected from N, O and S.The nitrogen and sulfur heteroatoms may optionally be oxidized and thenitrogen heteroatoms may optionally be quaternized. Examples ofheterocycle include, but are not limited to, tetrahydropyridyl,piperidinyl, morpholinyl, tetrahydrofuranyl, tetrahydrothienyl,piperazinyl, 1-azepanyl, imidazolinyl, 1,4-dioxanyl and the like.

As used herein, the terms “C₇-C₁₆ aralkyl” refer to an alkyl group asdefined herein that is substituted by one or more aryl groups as definedherein. Aralkyl groups include, for example, benzyl groups.

Various embodiments of the disclosure are described herein. It will berecognized that features specified in each embodiment may be combinedwith other specified features to provide further embodiments.

The present disclosure encompasses the compounds of formula (I), (II),(III), their tautomers, enantiomers, diastereomers, racemates ormixtures thereof, and their hydrates, solvates or pharmaceuticallyacceptable salts.

The terms “pharmaceutically acceptable salts” refers to salts thatretain the biological effectiveness and properties of the compounds ofthis disclosure and, which typically are not biologically or otherwiseundesirable.

Any formula given herein is also intended to represent unlabeled as wellas isotopically forms of the compounds, like deuterium labeled compoundsor ¹⁴C-labeled compounds.

Compound of Formula (I)

The present disclosure relates to a compound of formula (I)

-   -   wherein    -   R₁ and R₂ are independently selected from H, halogen, C₁-C₆        alkyl, C₃-C₆ cycloalkyl, C₁-C₆ haloalkyl, C₂-C₆ alkenyl, C₂-C₆        alkynyl, heterocyclyl having 5 to 10 ring atoms, aryl having 6        to 10 ring atoms, heteroaryl having 5 to 10 ring atoms, and        C₇-C₁₆ aralkyl, said alkyl, cycloalkyl, haloalkyl, alkenyl,        alkynyl, heterocyclyl, aryl, heteroaryl and aralkyl being        optionally substituted with one or more substituents        independently selected from oxo, halogen, C₁-C₆ alkyl, C₁-C₆        alkoxy, C₁-C₆ haloalkyl, —OH, —NR″R′″, —NO₂, —CN and —(CO)—R;    -   or R₁ and R₂, together with the carbon-carbon double bond        between them, form a 6 to 10 membered aryl or heteroaryl ring,        said aryl and heteroaryl being optionally substituted with one        or more substituents independently selected from halogen, C₁-C₆        alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —OH, —NR″R′″, —NO₂, —CN        and —(CO)—R;    -   Y is −O— or —S—;    -   R₃ is selected from C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₁-C₆        haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, heterocyclyl having 5        to 10 ring atoms, aryl having 6 to 10 ring atoms, heteroaryl        having 5 to 10 ring atoms and C₇-C₁₆ aralkyl, said alkyl,        cycloalkyl, haloalkyl, alkenyl, alkynyl, heterocyclyl, aryl,        heteroaryl and aralkyl being optionally substituted with one or        more substituents independently selected from oxo, halogen,        C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —OH, —NR″R″, —NO₂,        —CN and —(CO)—R;

each R is independently selected from H, C₁-C₆ alkyl, C₁-C₆ alkoxy and—NR″R′″;

each R″ and R′″ is independently selected from H and C₁-C₆ alkyl.

In one embodiment, the disclosure relates to a compound of formula (I)wherein

-   -   R₁ and R₂ are independently selected from H, halogen, C₁-C₆        alkyl, C₃-C₆ cycloalkyl, C₁-C₆ haloalkyl, heterocyclyl having 5        to 10 ring atoms, aryl having 6 to 10 ring atoms, heteroaryl        having 5 to 10 ring atoms and C₇-C₁₆ aralkyl, said alkyl,        cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl being        optionally substituted with one or more substituents        independently selected from halogen, C₁-C₆ alkyl, C₁-C₆ alkoxy,        C₁-C₆ haloalkyl, —NO₂, —CN and —(CO)—R;    -   or R₁ and R₂, together with the carbon-carbon double bond        between them, form a 6 membered aryl or heteroaryl ring, said        aryl and heteroaryl being optionally substituted with one or        more substituents independently selected from halogen, C₁-C₆        alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —NO₂, —CN and —(CO)—R;    -   Y is −O— or —S—;    -   R₃ is selected from C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₁-C₆        haloalkyl, heterocyclyl having 5 to 10 ring atoms, aryl having 6        to 10 ring atoms, heteroaryl having 5 to 10 ring atoms and        C₇-C₁₆ aralkyl, said alkyl, cycloalkyl, heterocyclyl, aryl,        heteroaryl and aralkyl being optionally substituted with one or        more substituents independently selected from halogen, C₁-C₆        alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —NO₂, —CN and —(CO)—R;    -   each R is independently selected from H, C₁-C₆ alkyl and C₁-C₆        alkoxy.

In one embodiment, R₁ is selected from H, C₁-C₆ alkyl and aryl having 6to 10 ring atoms, said alkyl, and aryl, being optionally substitutedwith one or more substituents independently selected from halogen, C₁-C₆alkyl, C₁-C₆ alkoxy, and —NO₂.

In another embodiment, R₂ is selected from H and C₁-C₆ alkyl, said alkylbeing optionally substituted with one or more substituents independentlyselected from halogen.

In another embodiment, R₁ and R₂ are independently selected from H,C₁-C₆ alkyl and aryl having 6 to 10 ring atoms, said aryl, beingoptionally substituted with one or more —NO₂.

In another embodiment, R₁ and R₂, together with the carbon-carbon doublebond between them, form a 6 membered aryl ring, optionally substitutedwith one or more substituents independently selected from halogen, C₁-C₆alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —NO₂, —CN and —(CO)—R; each R isindependently selected from H, C₁-C₆ alkyl and C₁-C₆ alkoxy.

In another embodiment, R₃ is selected from C₁-C₆ haloalkyl, heterocyclylhaving 5 to 10 ring atoms, aryl having 6 to 10 ring atoms and heteroarylhaving 5 to 10 ring atoms, said heterocyclyl, aryl, and heteroaryl beingoptionally substituted with one or more substituents independentlyselected from halogen, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —NO₂,—CN and —(CO)—R; each R being independently selected from C₁-C₆ alkyl.

In another embodiment, Y is —S—.

In another embodiment, the disclosure provides a compound of formula(I), wherein

-   -   R₁ and R₂, together with the carbon-carbon double bond between        them, form a 6-membered aryl ring;    -   Y is —O—;    -   R₃ is selected from aryl having 6 to 10 ring atoms, said aryl        being optionally substituted with one or more substituents        independently selected from halogen.

In another embodiment, the disclosure provides a compound of formula(I), wherein

-   -   R₁ and R₂ are independently selected from H, C₁-C₆ alkyl and        aryl having 6 to 10 ring atoms, said aryl, being optionally        substituted with one or more —NO₂;    -   or R₁ and R₂, together with the carbon-carbon double bond        between them, form a 6 membered aryl ring, optionally        substituted with one or more substituents independently selected        from halogen, C₁-C₆ alkyl and C₁-C₆ alkoxy,    -   Y is —S—;    -   R₃ is selected from C₁-C₆ haloalkyl, heterocyclyl having 5 to 10        ring atoms, aryl having 6 to 10 ring atoms, heteroaryl having 5        to 10 ring atoms, and C₇-C₁₆ aralkyl said heterocyclyl, aryl,        heteroaryl and aralkyl being optionally substituted with one or        more substituents independently selected from halogen, C₁-C₆        alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —NO₂, —CN and —(CO)—R;    -   each R is independently selected from C₁-C₆ alkyl.

In another embodiment, the disclosure provides a compound of formula(I), wherein

-   -   R₁ and R₂, together with the carbon-carbon double bond between        them, form a 6-membered aryl ring,    -   Y is —S—;    -   R₃ is selected from aryl having 6 to 10 ring atoms and        heteroaryl having 5 to 10 ring atoms, said aryl and heteroaryl        being optionally substituted with one or more substituents        independently selected from halogen, C₁-C₆ haloalkyl, C₁-C₆        alkoxy, —CN and —(CO)—R; each R being independently selected        from C₁-C₆ alkyl.

In another embodiment, the compound of formula (I) is selected from

According to one embodiment, the compound of formula (I) is selectedfrom

In a preferred embodiment, the compound of formula (I) is selected from

In some embodiments, the compounds of formula (I) as described hereinhave anti-proliferative activity towards melanoma cell lines.Accordingly, they may advantageously be used in a method for treatingcancer, and melanoma in particular.

Without being bound to this theory, the inventors hypothesized that, insome embodiments, the compounds of formula (I) induce the activation ofAMPK (AMP activated protein kinase) which is involved in the regulationof apoptosis, thereby inducing cell death to cancer cells.

Compounds of Formula (II) or (III)

Compounds of formula (I) wherein R₁ and R₂, together with thecarbon-carbon double bond between them, form an optionally substituted6-membered aryl ring, can spontaneously, or under specific conditionsdepending on the biguanide structure, rearrange to form compounds offormula (II) by opening of the benzothiazole or benzoxazole moiety andformation of a triazine ring. For example, in some cases, when acompound of formula (I) with Y is —S— is treated in oxidativeconditions, rearrangement to a compound of formula (II) occurs. When Y′is —SH, compounds of formula (II) can dimerize to form compounds offormula (III) by formation of an S—S bond.

Therefore, the disclosure also relates to a compound of formula (II) or(III)

-   -   wherein    -   Y′ is —SR₄ or −OR₅,    -   R₄ is selected from H, C₁-C₆ alkyl and protecting groups;    -   R₅ is selected from H and protecting groups;    -   each R′ is independently selected from halogen, C₁-C₆ alkyl,        C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —OH, —NR″R′″, —NO₂, —CN and        —(CO)—R;

n is 0 to 4;

-   -   R₃ is selected from C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₁-C₆        haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, heterocyclyl having 5        to 10 ring atoms, aryl having 6 to 10 ring atoms, heteroaryl        having 5 to 10 ring atoms and C₇-C₁₆ aralkyl, said alkyl,        cycloalkyl, haloalkyl, alkenyl, alkynyl, heterocyclyl, aryl,        heteroaryl and aralkyl being optionally substituted with one or        more substituents independently selected from oxo, halogen,        C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —OH, —NR″R′″, —NO₂,        —CN and —(CO)—R;    -   each R is independently selected from H, C₁-C₆ alkyl, C₁-C₆        alkoxy and —NR″R′″;    -   each R″ and R′″ is independently selected from H and C₁-C₆        alkyl.

In one embodiment, the disclosure relates to a compound of formula (II)or (III), wherein

-   -   Y′ is —SR₄ or −OR₅,    -   R₄ is selected from H, C₁-C₆ alkyl and protecting groups;    -   R₅ is selected from H and protecting groups;    -   each R′ is independently selected from halogen, C₁-C₆ alkyl,        C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —NO₂, —CN and —(CO)—R;    -   n is 0 to 4;    -   R₃ is selected from C₁-C₆ alkyl, C₁-C₆ haloalkyl, heterocyclyl        having 5 to 10 ring atoms, aryl having 6 to 10 ring atoms,        heteroaryl having 5 to 10 ring atoms and C₇-C₁₆ aralkyl, said        alkyl, heterocyclyl, aryl, heteroaryl and aralkyl being        optionally substituted with one or more substituents        independently selected from halogen, C₁-C₆ alkyl, C₁-C₆ alkoxy,        C₁-C₆ haloalkyl, —NO₂, —CN and —(CO)—R; each R is independently        selected from H, C₁-C₆ alkyl and C₁-C₆ alkoxy.

In one embodiment, n is 1 and R′ is selected from halogen and C₁-C₆alkoxy. In another embodiment, n is 1 and R′ is selected from halogen,C₁-C₆ alkyl, C₁-C₆ alkoxy, and —NO₂.

In another embodiment, n is 0.

In another embodiment, Y′ is —SR₄ and R₄ is selected from C₁-C₆ alkyland protecting groups, preferably, the protecting group is selected fromany group linked by a disulfide function, thioesters, alkyl, alkenyl andalkynyl thioethers, benzyl thioethers, alkylarylmethyl thioethers, andtriarylmethylthioethers.

In another embodiment, Y′ is —OR₅ and R₅ is selected from H andprotecting groups, preferably, the protecting group is selected fromesters, alkenyl and alkynyl ethers, silylated ethers, alkoxymethylethers, benzyl ethers tetrahydropyranyl ethers, pentoses, and hexoses.

In an embodiment, Y′ is —OH, n is 1, and R′ is selected from halogen,and —NO₂.

In another embodiment, R₃ is selected from C₁-C₆ haloalkyl, aryl having6 to 10 ring atoms, heteroaryl having 5 to 10 ring atoms and C₇-C₁₆aralkyl, said aryl, heteroaryl and aralkyl being optionally substitutedwith one or more substituents independently selected from halogen, C₁-C₆alkoxy, C₁-C₆ haloalkyl, and —CN.

In another embodiment, the disclosure provides a compound of formula(II) or (III), wherein

-   -   Y′ is —SR₄, R₄ being selected from C₁-C₆ alkyl;    -   each R′ is independently selected from halogen and C₁-C₆ alkoxy;    -   n is 0 to 1;    -   R₃ is selected from C₁-C₆ haloalkyl, aryl having 6 to 10 ring        atoms, heteroaryl having 5 to 10 ring atoms, and C₇-C₁₆ aralkyl,        said aryl, heteroaryl and aralkyl being optionally substituted        with one or more substituents independently selected from        halogen, C₁-C₆ alkoxy, and —CN.

In another embodiment, the disclosure provides a compound of formula(II), wherein

-   -   Y′ is —SR₄, R₄ being selected from C₁-C₆ alkyl;    -   each R′ is independently selected from halogen and C₁-C₆ alkoxy;    -   n is 0 to 1;    -   R₃ is selected from C₁-C₆ haloalkyl, aryl having 6 to 10 ring        atoms, heteroaryl having 5 to 10 ring atoms and C₇-C₁₆ aralkyl,        said aryl, heteroaryl and aralkyl being optionally substituted        with one or more substituents independently selected from        halogen, C₁-C₆ alkoxy, and —CN.

In another embodiment, the disclosure provides a compound of formula(II), wherein

-   -   Y′ is —OR₅ and R₅ is H;    -   each R′ is independently selected from halogen, C₁-C₆ alkyl, and        —NO₂,    -   n is 0 to 1;    -   R₃ is selected from C₁-C₆ haloalkyl, aryl having 6 to 10 ring        atoms, and heteroaryl having 5 to 10 ring atoms, said aryl, and        heteroaryl being optionally substituted with one or more        substituents independently selected from halogen, C₁-C₆ alkoxy,        and —CN.

In another embodiment, the disclosure provides a compound of formula(II), wherein

-   -   each R′ is independently selected from halogen and C₁-C₆ alkoxy;    -   n is 0 to 1;    -   R₃ is selected from C₁-C₆ haloalkyl, aryl having 6 to 10 ring        atoms, heteroaryl having 5 to 10 ring atoms and C₇-C₁₆ aralkyl,        said aryl, heteroaryl and aralkyl being optionally substituted        with one or more substituents independently selected from        halogen, C₁-C₆ alkoxy, and —CN.

In another embodiment, the disclosure provides a compound of formula(II) selected from

In another embodiment, the disclosure provides a compound of formula(II) selected from

In another embodiment, the disclosure provides a compound of formula(III), wherein

-   -   each R′ is independently selected from halogen and C₁-C₆ alkoxy;    -   n is 0 to 1;    -   R₃ is selected from C₁-C₆ haloalkyl, aryl having 6 to 10 ring        atoms, heterocyclyl having 5 to 10 ring atoms, heteroaryl having        5 to 10 ring atoms and C₇-C₁₆ aralkyl, said aryl, heteroaryl,        heterocycle, and aralkyl being optionally substituted with one        or more substituents independently selected from halogen, C₁-C₆        alkoxy, C₁-C₆ alkyl, —NR″R′″, —NO₂, —CN and —(CO)—R;    -   each R is independently selected from C₁-C₆ alkyl;    -   R″ and R′″ are H.

In another embodiment, the disclosure provides a compound of formula(III) selected from

In another embodiment, the disclosure provides a compound of formula(III) selected from

In a preferred embodiment, the disclosure provides a compound of formula(III) selected from

In compounds of formula (II), the hydroxyl or thiol function can beprotected with a protecting group. As used herein, protecting groupsrefers to any group cleavable in biological medium, particularly byhydrolysis or removal via the plasmatic enzymes or bioorganicnucleophiles such as glutathione. Suitable protecting group for hydroxylinclude [but are not limited to esters, alkyl, alkenyl and alkynylethers, silylated ethers, alkoxymethyl ethers, benzyl ethers,tetrahydropyranyl ethers, pentoses, hexoses. Suitable protecting groupfor thiol group include [but are not limited to any group linked by adisulfide function, thioesters, alkyl, alkenyl and alkynyl thioethers,benzyl thioethers, alkylarylmethyl thioethers, triarylmethylthioethers.

Compounds of formula (II) or (III) as described herein have a highbiological activity towards melanoma, including melanomas resistant toBRAF inhibitors. They can therefore be used in a method for treatingcancer, and melanoma in particular.

Without being bound to this theory, the inventors hypothesized thatcompounds of formula (II) and (III) are prodrugs of compounds of formula(I). After administration to a subject, the compounds of formula (II)and (III) are modified through in vivo physiological action, into acompound of formula (I).

Pharmaceutical Composition

The disclosure also relates to a pharmaceutical composition comprising acompound of formula (I), (II) or (III), and a pharmaceuticallyacceptable carrier.

“Pharmaceutically” or “pharmaceutically acceptable” refers to molecularentities and compositions that do not produce an adverse, allergic orother untoward reaction when administered to a mammal, especially ahuman, as appropriate. A pharmaceutically acceptable carrier orexcipient refers to a non-toxic solid, semi-solid or liquid filler,diluent, encapsulating material or formulation auxiliary of any type.

The form of the pharmaceutical compositions, the route ofadministration, the dosage and the regimen naturally depend upon thecondition to be treated, the severity of the illness, the age, weight,and sex of the patient, etc.

The pharmaceutical compositions of the disclosure can be formulated fora topical, oral, intranasal, intraocular, intravenous, intramuscular orsubcutaneous administration and the like.

The pharmaceutical compositions can take the form of tablets, pills,capsules, semisolids, powders, sustained release formulations,solutions, suspensions, emulsions, syrups, elixirs, aerosols, or anyother appropriate compositions; and comprise at least one compoundaccording this disclosure.

Because of their ease in administration, tablets and capsules representthe most advantageous oral dosage unit form, in which case solidpharmaceutical carriers are obviously employed. If desired, tablets maybe sugar coated or enteric coated by standard techniques. The tablets orpills can be coated to provide a dosage form affording the advantage ofprolonged action. For example, the tablet or pills can comprise an innerdosage and an outer dosage component, the latter being in the form of anenvelope over the former. The two components can be separated by anenteric layer, which serves to resist disintegration in the stomach andpermits the inner component to pass intact into the duodenum or to bedelayed in release. A variety of material can be used for such entericlayers or coatings, such materials including a number of polymeric acidswith such materials as shellac, cetyl alcohol and cellulose acetate.

The compound of the disclosure and the further agent may also beadministered in the form of liposome delivery systems, such as smallunilamellar vesicles, large unilamellar vesicles and multilamellarvesicles. Liposomes can be formed from a variety of phospholipids, suchas cholesterol, stearylamine or phosphatidylcholines.

Preferably, the pharmaceutical compositions contain vehicles which arepharmaceutically acceptable for a formulation capable of being injected.These may be in particular isotonic, sterile, saline solutions(monosodium or disodium phosphate, sodium, potassium, calcium ormagnesium chloride and the like or mixtures of such salts), or dry,especially freeze-dried compositions which upon addition, depending onthe case, of sterilized water or physiological saline, permit theconstitution of injectable solutions.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions or dispersions; formulations including sesame oil,peanut oil or aqueous propylene glycol; and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In all cases, the form must be sterile and must be fluid tothe extent that easy syringability exists. It must be stable under theconditions of manufacture and storage and must be preserved against thecontaminating action of microorganisms, such as bacteria and fungi.Solutions of the active compounds as free base or pharmacologicallyacceptable salts can be prepared in water suitably mixed with asurfactant, such as hydroxypropylcellulose. Dispersions can also beprepared in glycerol, liquid polyethylene glycols, mixtures thereof andin oils. Under ordinary conditions of storage and use, thesepreparations contain a preservative to prevent the growth ofmicroorganisms.

The carrier can also be a solvent or dispersion medium containing, forexample, water, ethanol, polyol (for example, glycerol, propyleneglycol, and liquid polyethylene glycol, and the like), suitable mixturesthereof, and vegetables oils. The proper fluidity can be maintained, forexample, by the use of a coating, such as lecithin, by the maintenanceof the required particle size in the case of dispersion and by the useof surfactants. The prevention of the action of microorganisms can bebrought about by various antibacterial and antifungal agents, forexample, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, andthe like. In many cases, it will be preferable to include isotonicagents, for example, sugars or sodium chloride. Prolonged absorption ofthe injectable compositions can be brought about by the use in thecompositions of agents delaying absorption, for example, aluminiummonostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the activecompounds in the required amount in the appropriate solvent with variousof the other ingredients enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the various sterilized active ingredients into a sterilevehicle which contains the basic dispersion medium and the requiredother ingredients from those enumerated above. In the case of sterilepowders for the preparation of sterile injectable solutions, thepreferred methods of preparation are vacuum-drying and freeze-dryingtechniques which yield a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof. Upon formulation, solutions will be administered in amanner compatible with the dosage formulation and in such amount as istherapeutically effective. The formulations are easily administered in avariety of dosage forms, such as the type of injectable solutionsdescribed above, but drug release capsules and the like can also beemployed.

For parenteral administration in an aqueous solution, for example, thesolution may be suitably buffered and the liquid diluent first renderedisotonic with sufficient saline or glucose. These particular aqueoussolutions are especially suitable for intravenous, intramuscular,subcutaneous and intraperitoneal administration. In this connection,sterile aqueous media which can be employed will be known to those ofskill in the art in light of the present disclosure. For example, onedosage could be dissolved in 1 ml of isotonic NaCl solution and eitheradded to 1000 ml of hypodermoclysis fluid or injected at the proposedsite of infusion, (see for example, “Remington's PharmaceuticalSciences” 15th Edition, pages 1035-1038 and 1570-1580). Some variationin dosage will necessarily occur depending on the condition of thesubject being treated. The person responsible for administration will,in any event, determine the appropriate dose for the individual subject.

For aerosol administration, the compound of the disclosure and thefurther agent are preferably supplied in finely divided from along witha surfactant and propellant. The surfactant must, of course, benontoxic, and preferably soluble in the propellant. Representative ofsuch agents are the esters or partial esters of fatty acids containingfrom 6 to 22 carbon atoms, such as caproic, octanoic lauric, palmitic,stearic, linoleic, linolenic, olesteric and oleic acids with analiphatic polyhydric alcohol or its cyclic anhydride. Mixed esters, suchas mixed or natural glycerides may be employed. A carrier can also beincluded, as desired, as with, e.g., lecithin for intranasal delivery.An example includes a solution in which each milliliter included 7.5 mgNaCl, 1.7 mg citric acid monohydrate, 3 mg disodium phosphate dihydrateand 0.2 mg benzalkonium chloride solution (50%) (Gozes et al., J MolNeurosci. 19(1-2):167-70 (2002)).

Suitable compositions for topical application include aqueous solutions,suspensions, ointments, creams, gels or sprayable formulations, e.g.aerosol administration.

The doses used for the administration can be adapted as a function ofvarious parameters, and in particular as a function of the mode ofadministration used, of the relevant pathology, or alternatively of thedesired duration of treatment. It will be appreciated that appropriatedosages of the compounds, and compositions comprising the compounds, canvary from patient to patient. Determining the optimal dosage willgenerally involve the balancing of the level of therapeutic benefitagainst any risk or deleterious side effects of the treatments describedherein. The selected dosage level will depend on a variety of factorsincluding, but not limited to, the activity of the particular compound,the route of administration, the time of administration, the rate ofexcretion of the compound, the duration of the treatment, other drugs,compounds, and/or materials used in combination, and the age, sex,weight, condition, general health, and prior medical history of thepatient. The amount of compound and route of administration willultimately be at the discretion of the physician, although generally thedosage will be to achieve local concentrations at the site of actionwhich achieve the desired effect without causing substantial harmful ordeleterious side-effects.

According to an embodiment, the pharmaceutical composition comprises anon-ionic emulsifier, preferably Kolliphor EL. The presence of suchnon-ionic emulsifier allows diminishing the final concentration of DMSOin the composition. The compound of formula (I), (II) or (III) can besolubilized in a non-ionic emulsifier, like Kolliphor EL.

Method of Use

The compounds of formula (I), (II) or (III) exhibit valuablepharmaceutical properties as indicated in the in vitro and in vivo testsprovided in the examples and are therefore indicated for therapy.

The disclosure also relates to a compound of formula (I), (II) or (III)for use as a medicament.

The disclosure also relates to a compound of formula (I), (II) or (III)for use in a method for treating cancer.

As used herein, the term “cancer” has its general meaning in the art andincludes an abnormal state or condition characterized by rapidlyproliferating cell growth. The term is meant to include all types ofcancerous growths or oncogenic processes, metastatic tissues ormalignantly transformed cells, tissues or organs, irrespective ofhistopathologic type or stage of invasiveness. The term cancer includesmalignancies of the various organ systems, such as affecting skin, lung,breast, thyroid, lymphoid, gastrointestinal, and genito-urinary tract,as well as adenocarcinomas which include malignancies such as most coloncancers, renal-cell carcinoma, prostate cancer and/or testicular tumors,non-small cell carcinoma of the lung, cancer of the small intestine andcancer of the oesophages.

Examples of cancer include, but are not limited, to hematologicalmalignancies such as B-cell lymphoid neoplasm, T-cell lymphoid neoplasm,non-hodgkin lymphoma (NHL), B-NHL, T-NHL, chronic lymphocytic leukemia(CLL), small lymphocytic lymphoma (SLL), mantle cell lymphoma (MCL),NK-cell lymphoid neoplasm, and myeloid cell lineage neoplasm. Examplesof non-hematological cancers include, but are not limited to, skincancer, colon cancer, breast cancer, lung cancer, brain cancer, prostatecancer, head and neck cancer, pancreatic cancer, bladder cancer,colorectal cancer, bone cancer, cervical cancer, liver cancer, oralcancer, esophageal cancer, thyroid cancer, kidney cancer, stomach cancerand testicular cancer.

In specific embodiment, the disclosure relates to a compound of formula(I), (II) or (III) for use in a method for treating melanoma. In oneparticular embodiment, the disclosure also relates to a compound offormula (I), (II) or (III) for use in a method for treating BRAFinhibitor-resistant melanoma.

In a specific embodiment, compounds of formula (I), (II) or (III) inducethe activation of AMPK. Metformin, a drug normally used to treat type IIdiabetes, also induces the activation of AMPK and it has been shown thatmetformin also inhibit the growth of melanoma cells. Therefore, byanalogy with metformin, in one particular embodiment, the disclosurealso relates to a compound of formula (I), (II) or (III) for use in amethod for treating type II diabetes.

The disclosure relates to a method for treating cancer, said methodcomprising administering to a subject a therapeutically efficient amountof

-   -   (i) a compound of formula (I),    -   (ii) a compound of formula (II) or (III), or    -   (iii) a pharmaceutical composition as described herein.

The terms “therapeutically efficient amount” of a compound refer to anamount of the compound that will elicit the biological or medicalresponse of a subject, for example, ameliorate the symptoms, alleviateconditions, slow or delay disease progression, or prevent a disease.

The disclosure also relates to the use of a compound of formula (I),(II) or (III), for the manufacture of a medicament for the treatment ofcancer. In one embodiment, cancer is melanoma. In one embodiment, canceris BRAF inhibitor-resistant melanoma.

FIGURES LEGENDS

In all the figures, the bars indicate the mean±SEM: *p<0.05; **p<0.01;***p<0.001.

FIG. 1A represents the cell viability of A375 S cells treated with 5 μMof CRO15 or PLX4032 at time indicated on the graph.

FIG. 1B represents the cell viability of A375 S cells treated withdifferent concentrations of CRO15 or with 5 μM PLX4032 for 48 hours.

FIG. 1C represents the viability of different melanoma cells withvarious mutations treated with 5 μM CRO15 for 48 hours. Mutations arespecified next to name of melanoma cell lines. Mutated proteins aresymbolized with “*”.

FIG. 1D represents the viability of patient melanoma cells treated with5 μM CRO15 for 48 hours. Mutated proteins are symbolized with “*”

FIG. 1E represents the viability of normal cells treated with 5 μM CRO15for 48 hours.

FIG. 2 represents the results of the western blot assays.

FIG. 3A represents the evolution of the tumoral volume of xenograft miceinoculated subcutaneously with A375 sensitive melanoma cells and treatedwith CRO15 or PLX4032.

FIG. 3B represents the tumoral weight of xenograft mice inoculatedsubcutaneously with A375 sensitive melanoma cells and treated with CRO15or PLX4032, after mice euthanasia.

FIG. 4A represents the viability of both sensitive and resistant A375melanoma cells treated with CRO15 or PLX4032.

FIG. 4B represents the evolution of the tumoral volume of xenograft miceinoculated subcutaneously with A375 resistant melanoma cells and treatedwith CRO15 or PLX4032.

FIG. 4C represents the tumoral weight of xenograft mice inoculatedsubcutaneously with A375 resistant melanoma cells and treated with CRO15or PLX4032, after mice euthanasia.

FIG. 5 represents the cell viability of DR6 cells treated with differentconcentrations of CRO15 or with 1 μM of vemurafenib and 0.5 μM ofcobimetinib for 24 hours.

FIG. 6A represents the evolution of the tumoral volume of xenograft miceinoculated subcutaneously with A375 resistant melanoma cells and treatedwith MTF319 or PLX4032.

FIG. 6B represents the tumoral weight of xenograft mice inoculatedsubcutaneously with A375 resistant melanoma cells and treated withMTF319 or PLX4032, after mice euthanasia.

FIG. 7A represents the evolution of the tumoral volume of allograft miceinoculated subcutaneously with murine melanoma cells and treated withCRO15.

FIG. 7B represents the cell viability of WM9 cells after treatment for 8weeks with PLX4032, CRO15 or MTF255 and stimulation with 10 μM of eachdrug for 48 h.

EXAMPLES Experimental Procedures

Chemical Synthesis and Characterization

Methanol, ethyl acetate, diethyl ether and dichloromethane werepurchased from Carlo Erba, and used as received. Anhydrous DMF (99.8%stored under septum) was purchased from Sigma Aldrich, and used asreceived. All chemicals were purchased from Aldrich, Fisher or AlfaAesar and used without further purification. Thin layer chromatography(TLC) was performed on precoated Merck 60 GF254 silica gel plates andrevealed first by visualization under UV light (254 nm and 360 nm) ¹Hand ¹³C NMR spectra were recorded on a Bruker Advance 200 MHzspectrometer or a Bruker Advance 400 MHz or a Bruker Advance 500 MHz.Mass spectra (ESI-MS) were recorded on a Bruker (Daltonics Esquire3000+). HRMS spectra were recorded on a ThermoFisher Q Exactive (ESI-MS)at a resolution of 140 000 at m/z 200. The purity of compounds wasfurther assayed by HPLC analysis on a JASCO PU-2089 apparatus withSupelco analytical column Ascentis Express C18, 100 mm×46 mm 5 μM.Eluent A: water with 1‰ formic acid. Eluent B: CH₃CN with 1‰ formicacid. Method 1: 30% B for 1 min, 30% B to 100% B over 5 min, 100% B for2.5 min then from 100% B to 30% B over 30 sec, 30% B for 7 min (16 minin total). Method 2: 30% B for 1 min, 30% B to 100% B over 5 min, 100% Bfor 20 min then from 100% B to 30% B over 1 min, 30% B for 4 min (31 minin total). Method 3: 30% B for 1 min, 30% B to 100% B over 5 min, 100% Bfor 2.5 min then from 100% B to 30% B over 30 sec (9 min in total).Method 4: 10% B for 10 min: 10% B to 95% B over 8 min, 95% B for 2 min,then from 95% B to 10% B over 4 min, 10% B for 1 min (25 min in total).

Synthetic Procedures and Characterizations:

General Procedure (A) for the Formation of Biguanides of Formula I

To a solution of the corresponding guanidine (1 equiv) inN,N-dimethylformamide (25 mL/g of guanidine) was added sodium hydride(60% dispersion in mineral oil, 1.5 eq.) and the mixture was stirred for30 min at r.t. To this solution was added the corresponding nitrile (1equiv) in one portion. The reaction was stirred overnight at roomtemperature and monitored by TLC. After completion of the reaction, themixture was poured into water (200 mL/g of guanidine) and theprecipitate was collected and washed with water, methanol and diethylether.

General Procedure (B) for the Formation ofN2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-aryl/hetaryl-1,3,5-triazine-2,4-diamines)of formula II and III (B)

A solution of the corresponding biguanide (1 equiv) in technical gradeethanol (approx. 25 mL/100 mg of biguanide) was stirred at refluxtemperature and monitored by LCMS. After full conversion (approx 6-7hours), the precipitate formed was filtered and washed with technicalgrade ethanol.

General Procedure (C) for the formation ofN2,N2′-(disulfanediylbis(4-alkyl/hal-2,1-phenylene))bis(6-(trichloromethyl)-1,3,5-triazine-2,4-diamines)of Formula II and III

To a solution of the corresponding guanidine (1 equiv) in technicalgrade ethanol (approx. 10 mL/g of guanidine), trichloroacetonitrile (2equiv) was added and the reaction mixture was stirred at refluxtemperature and monitored by LCMS. After full conversion (approx 6-7hours), the precipitate formed was filtered and washed with technicalgrade ethanol and diethyl ether.

General Procedure (D) for the formation of2-((6-imino-4-(trichloromethyl)-1,6-dihydro-1,3,5-triazin-2-yl)amino)phenols

To a solution of corresponding guanidine (1 equiv.) in technical gradeethanol (25 mL/g of guanidine) trichloroacetonitrile (10 equiv.) wasadded and the reaction mixture was stirred in a sealed tube under argonatmosphere at 60° C. After full conversion (approx. 18 hours), themixture was concentrated to dryness. The residue was purified by silicagel flash chromatography.

General Procedure (E) for the formation of2-(4-amino-6-phenyl-1,3,5-triazin-2-yl)amino)phenols

To a solution of corresponding guanidine (1 equiv.) inN,N-dimethylformamide (25 mL/g of guanidine) sodium hydride (60%dispersion in mineral oil, 1.1 equiv.) was added and the reactionmixture was stirred in a sealed tube under argon atmosphere. When thegas evolution stopped to this solution was added corresponding nitrile(1 equiv.) and the tube was sealed. The resulting solution was thenstirred at 80° C. After full conversion (approx. 18 hours), the mixturewas concentrated to dryness. The residue was purified by silica gelflash chromatography.

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(trichloromethyl)-1,3,5-triazine-2,4-diamine)(CRO15)

A solution of 1-(benzo[d]thiazol-2-yl)guanidine (10.0 g, 52 mmol) andtrichloroacetonitrile (10.0 mL, 1.92 mmol) in technical grade ethanol(100 mL) was stirred at 75° C. After 1 h, a large amount of whiteprecipitate appeared in the yellow solution. After reaction completion(TLC monitoring, about 3 h), the suspension was cold down to r.t. andfiltered. The precipitate was washed with little amount of cold ethanoland dried at air. Recrystallization from acetone/diethyl ether affordedthe desired compound as a white solid (10.03 g, 57.5%). TLC: R_(f)(Et₂O/PE, 1/1, v/v)=0.23. ¹H NMR (200 MHz, Acetone-d6): δ 8.54 (s, 1H),7.96 (d, J=8.0 Hz, 1H), 7.58 (dd, J=7.8, 1.6 Hz, 1H), 7.35 (td, J=7.8,1.6 Hz, 1H), 7.22-6.93 (m, 3H). ¹³C NMR (101 MHz, Acetone-d6): δ 174.00,168.98, 166.37, 139.05, 133.76, 130.58, 129.92, 126.33, 125.34, 97.43.HRMS-ESI (m/z): [M+H]⁺ calc. for C₂₀H₁₅Cl₆N₁₀S₂ ⁺, 668.90482; Found:668.90497. HPLC (λ₂₈₀): Purity 97.4%; t_(R): 7.958 min (method 2).

N2,N2′-(disulfanediylbis(4-methoxy-2,1-phenylene))bis(6-(trichloromethyl)-1,3,5-triazine-2,4-diamine)(MTF-232)

Synthesized following the general procedure C using1-(6-methoxybenzo[d]thiazol-2-yl)guanidine (300 mg, 1.35 mmol) to affordthe titled compound as a green powder (418 mg, 85%). ¹H NMR (400 MHz,DMSO-d6): δ 9.64 (s, 1H), 7.59 (s, 1H), 7.45 (s, 1H), 7.20 (d, J=8.7 Hz,1H), 7.10 (s, 1H), 6.84 (dd, J=8.7, 2.8 Hz, 1H), 3.70 (s, 3H). ¹³C NMR(50 MHz, DMSO-d6): δ 172.28, 167.38, 166.07, 158.26, 135.44, 129.22,128.28, 113.08, 112.20, 96.77, 55.48. HPLC (λ₂₈₀): Purity 100.0%; t_(R):7.433 min (method 3).

N2,N2′-(disulfanediylbis(4-chloro-2,1-phenylene))bis(6-(trichloromethyl)-1,3,5-triazine-2,4-diamine)(MTF-233)

Synthesized following the general procedure C using1-(6-chlorobenzo[d]thiazol-2-yl)guanidine (300 mg, 1.32 mmol) to affordthe titled compound as a white powder (454 mg, 93%). ¹H NMR (400 MHz,DMSO-d6): δ 9.66 (s, 1H), 7.71 (s, 1H), 7.58 (s, 2H), 7.40 (d, J=8.5 Hz,1H), 7.35 (dd, J=8.5, 2.3 Hz, 1H). ¹³C NMR (50 MHz, DMSO-d6): δ 172.39,167.34, 165.59, 135.71, 134.89, 131.29, 129.12, 127.91, 127.52, 96.63.HPLC (λ₂₈₀): Purity 95.1%; t_(R): 13.767 min (method 1).

N2,N2′-(disulfanediylbis(4-fluoro-2,1-phenylene))bis(6-(trichloromethyl)-1,3,5-triazine-2,4-diamine)(MTF-234)

Synthesized following the general procedure C using1-(6-fluorobenzo[d]thiazol-2-yl)guanidine (300 mg, 1.42 mmol) to affordthe titled compound as a white powder (457 mg, 91%). ¹H NMR (200 MHz,DMSO-d6): δ 9.06 (br. s., 2H), 7.69 (br. s., 1H), 7.57 (br. s., 1H),7.45-7.30 (m, 2H), 7.14 (t, 1H, J=8.6 Hz, H1). ¹³C NMR (50 MHz,DMSO-d6): δ 172.42, 167.38, 165.94, 160.87 (d), 136.40, 131.93, 130.04,114.76 (d), 113.61 (d), 96.69. HPLC (λ₂₈₀): Purity 98.7%; t_(R): 6.858min (method 3).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-bromobenzimidamide (MTF-242)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and3-bromobenzonitrile (0.95 g, 5.20 mmol) to afford the titled compound asa white-yellowish powder (708 mg, 36%). ¹H NMR (500 MHz, DMSO-d6): δ10.29 (br. s, 1H), 9.36 (br. s, 1H), 8.94 (br. s, 1H), 8.26 (s, 1H),8.11 (s, 1H), 8.03 (d, J=7.9 Hz, 1H), 7.80 (d, J=7.7 Hz, 1H), 7.76 (dd,J=7.9, 1.1 Hz, 1H), 7.66 (d, J=8.0 Hz, 1H), 7.47 (t, J=7.9 Hz, 1H),7.39-7.30 (m, 1H), 7.24-7.15 (m, 1H). ¹³C NMR (50 MHz, DMSO-d6): δ172.23, 161.94, 160.52, 151.47, 137.42, 134.08, 131.23, 130.43, 126.51,125.66, 122.86, 121.70, 121.15, 119.78. HRMS-ESI (m/z): [M+H]⁺ calc. forC₁₅H₁₂BrN₅S⁺, 374.00696; Found: 374.00797. HPLC (λ₂₈₀): Purity 99.3%;t_(R): 6.775 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-chlorobenzimidamide(MTF-243)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and3-chlorobenzonitrile (715 mg, 5.20 mmol) to afford the titled compoundas a yellow powder (206 mg, 12%). ¹H NMR (500 MHz, DMSO-d6): δ 10.30(br. s, 1H), 9.37 (br. s, 1H), 8.90 (br. s, 1H), 8.12 (t, br. s, J=1.8Hz, 2H), 7.99 (d, J=7.9 Hz, 1H), 7.80 (d, J=7.7 Hz, 1H), 7.66 (d, J=7.9Hz, 1H), 7.65-7.60 (m, 1H), 7.54 (t, J=7.9 Hz, 1H), 7.38-7.30 (m, 1H),7.22-7.17 (m, 1H). ¹³C NMR (50 MHz, DMSO-d6): δ 172.22, 161.95, 160.57,151.47, 137.26, 133.22, 131.23, 131.20, 130.20, 127.53, 126.14, 125.67,122.87, 121.16, 119.79. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₅H₁₂ClN₅S⁺,330,05747; Found: 330,05774. HPLC (λ₂₈₀): Purity 99.2%; t_(R): 6.792 min(method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2-chlorobenzimidamide(MTF-244)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and2-chlorobenzonitrile (715 mg, 5.20 mmol) to afford the titled compoundas a white powder (326 mg, 19%). ¹H NMR (500 MHz, DMSO-d6): δ 10.14 (br.s, 1H), 9.30 (br. s, 1H), 8.81 (br. s, 1H), 8.09 (br. s, 1H), 7.80 (d,J=7.5 Hz, 1H), 7.65 (d, J=7.7 Hz, 1H), 7.53 (br. s, 2H), 7.50-7.40 (m,2H), 7.34 (t, J=7.1 Hz, 1H), 7.20 (t, J=7.1 Hz, 1H). ¹³C NMR (50 MHz,DMSO-d6): δ 172.40, 162.90, 162.00, 151.45, 136.67, 131.23, 130.66,130.29, 129.56, 129.46, 126.97, 125.63, 122.82, 121.16, 119.74. HRMS-ESI(m/z): [M+H]⁺ calc. for C₁₅H₁₂CN₅S⁺, 330.05747; Found: 330.05783. HPLC(λ₂₈₀): Purity 96.6%; t_(R): 6.500 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-chlorobenzimidamide(MTF-245)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and4-chlorobenzonitrile (715 mg, 5.20 mmol) to afford the titled compoundas a white-yellowish powder (721 mg, 42%). ¹H NMR (500 MHz, DMSO-d6): δ10.27 (br. s, 1H), 9.36 (br. s, 1H), 8.85 (br. s, 1H), 8.06 (d, br.s,J=8.6 Hz, 3H), 7.79 (d, J=7.4 Hz, 1H), 7.66 (d, J=7.9 Hz, 1H), 7.59 (d,J=8.6 Hz, 2H), 7.34 (t, J=7.7 Hz, 1H), 7.19 (t, J=7.6 Hz, 1H). ¹³C NMR(50 MHz, DMSO-d6): δ 172.22, 162.00, 160.95, 151.47, 136.28, 134.00,131.21, 129.44, 128.36, 125.65, 122.83, 121.15, 119.74. HRMS-ESI (m/z):[M+H]⁺ calc. for C₁₅H₁₂ClN₅S⁺, 330.05747; Found: 330.05743. HPLC (λ₂₈₀):Purity 98.8%; t_(R): 6.800 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-methylbenzimidamide(MTF-246)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) andp-tolunitrile (609 mg, 5.20 mmol) to afford the titled compound as abeige powder (145 mg, 9%). ¹H NMR (500 MHz, DMSO-d6): δ 10.28 (br. s,1H), 9.34 (br. s, 1H), 8.74 (br. s, 1H), 7.96 (d, br. s, J=7.9 Hz, 3H),7.79 (d, J=7.7 Hz, 1H), 7.65 (d, J=7.9 Hz, 1H), 7.33 (dd, J=18.6, 7.7Hz, 3H), 7.19 (t, J=7.5 Hz, 1H), 2.37 (s, 3H). ¹³C NMR (50 MHz,DMSO-d6): δ 172.21, 161.99, 161.33, 151.46, 137.15, 134.68, 131.21,129.49, 125.64, 122.82, 121.13, 119.74, 99.13. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₆H₁₅N₅S⁺, 310.11209; Found: 310.11218. HPLC (λ₂₈₀): Purity97.9%; t_(R): 6.592 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-iodobenzimidamide (MTF-247)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and4-iodobenzonitrile (1.19 g, 5.20 mmol) to afford the titled compound asa beige powder (832 mg, 38%). ¹H NMR (500 MHz, DMSO-d6): δ 10.24 (br. s,1H), 9.34 (br. s, 1H), 8.82 (br. s, 1H), 8.03 (br. s, 1H), 7.90 (d,J=8.5 Hz, 2H), 7.82 (d, J=8.5 Hz, 2H), 7.79 (d, J=7.8 Hz, 1H), 7.65 (d,J=7.9 Hz, 1H), 7.34 (t, J=7.6 Hz, 1H), 7.19 (t, J=8.0 Hz, 1H). ¹³C NMR(126 MHz, DMSO-d6): δ 172.20, 161.97, 161.31, 151.45, 137.13 (2C),134.68, 131.20, 129.47 (2C), 125.62, 122.79, 121.11, 119.72, 99.09.HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₅H₁₂IN₅S⁺, 421.99309; Found:421.99316. HPLC (λ₂₈₀): Purity 98.8%; t_(R): 6.900 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)benzimidamide (MTF-248)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) andbenzonitrile (0.54 mL, 5.20 mmol) to afford the titled compound as abeige powder (430 mg, 28%). ¹H NMR (500 MHz, DMSO-d6): δ 10.28 (br. s,1H), 9.35 (br. s, 1H), 8.80 (br. s, 1H), 8.03 (d, br. s, J=7.3 Hz, 3H),7.79 (d, J=7.6 Hz, 1H), 7.66 (d, J=7.9 Hz, 1H), 7.60-7.54 (m, 1H), 7.51(t, J=7.2 Hz, 2H), 7.34 (t, J=7.4 Hz, 1H), 7.19 (t, J=7.4 Hz, 1H). ¹³CNMR (50 MHz, DMSO-d6): δ 172.37, 162.29, 162.21, 151.57, 135.28, 131.44,131.26, 128.28, 127.60, 125.67, 122.82, 121.15, 119.75. HRMS-ESI (m/z):[M+H]⁺ calc. for C₁₅H₁₃N₅S⁺, 296.09644; Found: 296.09659. HPLC (λ₂₈₀):Purity 97.9%; t_(R): 6.358 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2-fluorobenzimidamide(MTF-249)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and2-fluorobenzonitrile (0.56 mL, 5.20 mmol) to afford the titled compoundas a red powder (179 mg, 11%). ¹H NMR (400 MHz, DMSO-d6): δ 10.16 (s,1H), 9.31 (s, 1H), 8.69 (s, 1H), 8.05 (s, 1H), 7.80 (d, J=7.7 Hz, 1H),7.72 (td, J=7.6, 1.4 Hz, 1H), 7.65 (d, J=7.9 Hz, 1H), 7.55 (ddd, J=9.4,7.3, 1.6 Hz, 1H), 7.37-7.28 (m, 3H), 7.20 (t, J=7.5 Hz, 1H). ¹³C NMR(101 MHz, DMSO-d6): δ 172.30 (s), 161.95 (s), 159.34 (d, J=250.6 Hz),160.01 (s), 151.43 (s), 132.26 (d, J=8.6 Hz), 131.21 (s), 130.39 (d,J=2.6 Hz), 125.62 (s), 124.59 (d, J=13.0 Hz), 124.28 (d, J=3.4 Hz),122.82 (s), 121.14 (s), 119.73 (s), 116.09 (d, J=21.9 Hz). ¹⁹F NMR (377MHz, DMSO-d6): δ −114.77. HPLC (λ₂₈₀): Purity 97.4%; t_(R): 6.792 min(method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2-bromobenzimidamide (MTF-250)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and2-bromobenzonitrile (946 mg, 5.20 mmol) to afford the titled compound asa beige powder (136 mg, 7%). ¹H NMR (400 MHz, DMSO-d6): δ 9.26 (s, 1H),7.67 (d, J=7.9 Hz, 1H), 7.54 (d, br. s, J=7.0 Hz, 2H), 7.44 (t, J=7.3Hz, 2H), 7.35 (t, J=7.1 Hz, 1H), 7.28-7.04 (m, 4H). ¹³C NMR (101 MHz,DMSO-d6): δ 173.15, 166.81, 165.00, 139.47, 136.49, 133.16, 132.85,130.48, 130.28, 128.84, 127.32, 127.03, 126.48, 126.12, 120.08. HRMS-ESI(m/z): [M+H]⁺ calc. for C₁₅H₁₂BrN₅S⁺, 374.00696; Found: 374.00702. HPLC(λ₂₈₀): Purity 97.8%; t_(R): 4.158 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)picolinimidamide (MTF-251)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and2-pyridinecarbonitrile (541 mg, 5.20 mmol) to afford the titled compoundas a white-yellowish powder (955 mg, 62%). ¹H NMR (200 MHz, DMSO-d6): δ10.10 (br. s, 1H), 9.40 (br. s, 1H), 8.92 (br. s, 1H), 8.72 (ddd, J=4.7,1.6, 0.9 Hz, 1H), 8.37 (dt, J=7.9, 1.0 Hz, 1H), 8.03 (br. s, td, J=7.7,1.7 Hz, 2H), 7.81 (dd, J=7.8, 0.8 Hz, 1H), 7.73-7.58 (m, 2H), 7.35 (td,J=7.7, 1.4 Hz, 1H), 7.20 (td, J=7.6, 1.2 Hz, 1H). ¹³C NMR (101 MHz,DMSO-d6): δ 170.26, 167.51, 165.72, 154.24, 149.29, 136.70, 133.47,128.60, 127.35, 126.57, 126.22, 125.45, 123.31. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₄H₁₂N₆S⁺, 291.09169; Found: 291.09174. HPLC (λ₂₈₀): Purity98.6%; t_(R): 6.075 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)nicotinimidamide (MTF-252)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol),3-pyridinecarbonitrile (541 mg, 5.20 mmol) to afford the titled compoundas a white-yellowish powder (1.49 g, 97%). ¹H NMR (200 MHz, DMSO-d6): δ10.20 (br. s, 1H), 9.34 (br. s, 1H), 9.17 (dd, J=2.2, 0.6 Hz, 1H), 8.95(br. s, 1H), 8.74 (dd, J=4.8, 1.6 Hz, 1H), 8.37-8.28 (m, 1H), 8.11 (br.s, 1H), 7.66 (dd, J=8.0, 0.6 Hz, 1H), 7.55 (ddd, J=8.0, 4.8, 0.8 Hz,1H), 7.35 (td, J=7.7, 1.4 Hz, 1H), 7.20 (td, J=7.6, 1.2 Hz, 1H). ¹³C NMR(101 MHz, DMSO-d6): δ 172.18, 161.92, 160.34, 151.88, 151.43, 148.75,135.10, 131.24, 130.86, 125.61, 123.24, 122.82, 121.11, 19.75. HRMS-ESI(m/z): [M+H]⁺ calc. for C₁₄H₁₂N₆S⁺, 297.09169; Found: 297.09174. HPLC(λ₂₈₀): Purity 100.0%; t_(R): 5.592 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)isonicotinimidamide (MTF-253)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and4-pyridinecarbonitrile (541 mg, 5.20 mmol) to afford the titled compoundas a white-yellowish powder (1.12 g, 73%). ¹H NMR (200 MHz, DMSO-d6): δ10.15 (br. s, 1H), 9.35 (br. s, 1H), 8.96 (br. s, 1H), 8.76 (dd, J=4.6,1.5 Hz, 2H), 8.15 (br. s, 1H), 7.91 (dd, J=4.6, 1.5 Hz, 2H), 7.81 (dd,J=7.7, 0.7 Hz, 1H), 7.67 (d, J=8.0 Hz, 1H), 7.42-7.30 (m, 1H), 7.27-7.14(m, 1H). ¹³C NMR (101 MHz, DMSO-d6): δ 172.12, 161.90, 160.07, 151.38,150.07, 142.53, 131.28, 125.64, 122.88, 121.43, 121.13, 119.81. HRMS-ESI(m/z): [M+H]⁺ calc. for C₁₄H₁₂N₆S⁺, 297.09169; Found: 297.09177. HPLC(λ₂₈₀): Purity 95.9%; t_(R): 5.308 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-methoxybenzimidamide(MTF-254)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and4-methoxybenzonitrile (692 mg, 5.20 mmol) to afford the titled compoundas a beige powder (135 mg, 8%). ¹H NMR (500 MHz, DMSO-d6): δ 10.27 (br.s, 1H), 9.30 (br. s, 1H), 8.69 (br. s, 1H), 8.03 (d, J=5.9 Hz, 2H), 7.92(br. s, 1H), 7.78 (d, J=5.4 Hz, 1H), 7.64 (d, J=5.8 Hz, 1H), 7.33 (s,1H), 7.18 (s, 1H), 7.05 (d, J=5.6 Hz, 2H), 3.83 (s, 3H). ¹³C NMR (50MHz, DMSO-d6): δ 172.33, 162.14, 161.96, 161.71, 151.58, 131.16, 129.40,127.19, 125.62, 122.72, 121.10, 119.64, 113.56, 55.41. HRMS-ESI (m/z):[M+H]⁺ calc. for C₁₆H₁₅N₅OS⁺, 326.10701; Found: 326.10718. HPLC (λ₂₈₀):Purity 95.3%; t_(R): 6.592 min (method 1).

3-acetyl-N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)benzimidamide(MTF-255)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and3-acetylbenzonitrile (755 mg, 5.20 mmol) to afford the titled compoundas a red powder (228 mg, 13%). ¹H NMR (200 MHz, DMSO-d6): δ 10.25 (br.s, 1H), 9.33 (br. s, 1H), 8.98 (br. s, 1H), 8.58 (t, J=1.6 Hz, 1H),8.30-8.22 (m, 1H), 8.20-7.97 (m, 2H), 7.81 (dd, J=7.8, 0.8 Hz, 1H), 7.68(t, J=7.7 Hz, 2H), 7.35 (td, J=7.7, 1.4 Hz, 1H), 7.20 (td, J=7.6, 1.2Hz, 1H), 2.66 (s, 3H). ¹³C NMR (50 MHz, DMSO-d6): δ 197.59, 172.28,162.07, 161.35, 151.50, 136.86, 135.70, 132.04, 131.24, 131.07, 128.79,127.31, 125.67, 122.85, 121.16, 119.77, 26.89. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₇H₁₅N₅OS⁺, 338.10701; Found: 338.10701. HPLC (λ₂₈₀): Purity95.2%; t_(R): 6.283 min (method 1).

3-bromo-N—(N-(4-phenylthiazol-2-yl)carbamimidoyl)benzimidamide (MTF-256)

Synthesized following the general procedure A using1-(4-phenylthiazol-2-yl)guanidine (1.00 g, 4.58 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 275 mg, 6.87 mmol) and3-bromobenzonitrile (833 mg, 4.58 mmol) to afford the titled compound asa yellow powder (715 mg, 39%). ¹H NMR (500 MHz, DMSO-d6): δ 10.36 (br.s, 1H), 9.15 (br. s, 1H), 8.79 (br. s, 1H), 8.25 (s, 1H), 8.02 (d, J=7.8Hz, 1H), 7.88 (d, br. s, J=7.3 Hz, 3H), 7.76 (dd, J=7.9, 1.0 Hz, 1H),7.47 (dd, J=9.1, 6.6 Hz, 2H), 7.43 (t, J=7.7 Hz, 2H), 7.31 (t, J=7.3 Hz,1H). ¹³C NMR (50 MHz, DMSO-d6): δ 173.11, 160.69, 160.04, 149.88,137.62, 134.52, 133.94, 130.38, 128.71, 127.61, 126.46, 125.61, 121.73,106.53. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₇H₁₄BrN₅S⁺, 400.02261; Found:400.02213. HPLC (λ₂₈₀): Purity 98.7%; t_(R): 7.033 min (method 1).

3-bromo-N—(N-(4-(3-nitrophenyl)thiazol-2-yl)carbamimidoyl)benzimidamide(MTF-257)

Synthesized following the general procedure A using1-(4-(3-nitrophenyl)thiazol-2-yl)guanidine (1.00 g, 3.80 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 228 mg, 5.70 mmol) and3-bromobenzonitrile (692 mg, 3.80 mmol) to afford the titled compound asa yellow powder (711 mg, 42%). ¹H NMR (500 MHz, DMSO-d6): δ 10.21 (s,1H), 8.78 (s, 2H), 8.62 (s, 1H), 8.34 (d, J=7.5 Hz, 1H), 8.23 (s, 1H),8.19 (br. s, 1H), 8.15 (d, J=7.8 Hz, 1H), 8.01 (d, J=7.5 Hz, 1H), 7.80(s, 1H), 7.76 (d, J=7.7 Hz, 1H), 7.72 (t, J=7.9 Hz, 1H), 7.48 (t, J=7.8Hz, 1H). ¹³C NMR (50 MHz, DMSO-d6): δ 173.32, 160.75, 159.98, 148.31,147.48, 137.51, 136.01, 133.95, 131.81, 130.37, 130.20, 126.47, 121.99,121.66, 119.77, 109.28. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₇H₁₃BrN₆O₂S⁺,445.00768; Found: 445.00806. HPLC (λ₂₈₀): Purity 99.1%; t_(R): 7.100 min(method 1).

N—(N-(4-(3-nitrophenyl)thiazol-2-yl)carbamimidoyl)picolinimidamide(MTF-259)

Synthesized following the general procedure A using1-(4-(3-nitrophenyl)thiazol-2-yl)guanidine (1.00 g, 3.80 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 228 mg, 5.70 mmol) and2-pyridinecarbonitrile (366 mL, 3.80 mmol) to afford the titled compoundas a yellow powder (796 mg, 57%). ¹H NMR (500 MHz, DMSO-d6): δ 10.03(br. s, 1H), 9.08 (br. s, 1H), 8.78 (br. s, 1H), 8.71 (d, J=4.2 Hz, 1H),8.66-8.61 (m, 1H), 8.38 (d, J=7.9 Hz, 1H), 8.35 (d, J=7.9 Hz, 1H), 8.16(dd, J=8.1, 1.5 Hz, 1H), 8.02 (td, J=7.8, 1.6 Hz, 1H), 7.91 (br. s, 1H),7.82 (s, 1H), 7.73 (t, J=8.0 Hz, 1H), 7.63 (ddd, J=7.4, 4.8, 1.0 Hz,1H). ¹³C NMR (50 MHz, DMSO-d6): δ 173.49, 161.02, 158.37, 150.67,148.58, 148.29, 147.51, 137.39, 135.99, 131.78, 130.16, 126.39, 122.10,121.99, 119.78, 109.31. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₆H₁₃N₇O₂S⁺,368.09242; Found: 368.09274. HPLC (λ₂₈₀): Purity 99.2%; t_(R): 6.850 min(method 1).

N—(N-(4-(3-nitrophenyl)thiazol-2-yl)carbamimidoyl)nicotinimidamide(MTF-260)

Synthesized following the general procedure A using1-(4-(3-nitrophenyl)thiazol-2-yl)guanidine (1.00 g, 3.80 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 228 mg, 5.70 mmol) and3-pyridinecarbonitrile (395 mg, 3.80 mmol) to afford the titled compoundas a yellow powder (656 mg, 47%). ¹H NMR (500 MHz, DMSO-d6): δ 10.14 (brs, 1H), 8.84 (br. s, 2H), 8.75 (d, J=5.8 Hz, 2H), 8.63 (s, 1H), 8.34 (d,J=7.8 Hz, 1H), 8.15 (dd, J=8.0, 1.4 Hz, 1H), 7.91 (d, J=5.8 Hz, 2H),7.86 (br. s, 1H), 7.82 (s, 1H), 7.72 (t, J=8.0 Hz, 1H). ¹³C NMR (50 MHz,DMSO-d6): δ 173.23, 160.74, 159.55, 150.07, 148.33, 147.49, 142.64,136.00, 131.80, 130.20, 122.01, 121.49, 119.79, 109.48. HRMS-ESI (m/z):[M+H]⁺ calc. for C₁₆H₁₃N₇O₂S⁺, 368.09242; Found: 368.09283. HPLC (λ₂₈₀):Purity 95.1%; t_(R): 6.350 min (method 1).

N—(N-(4-(3-nitrophenyl)thiazol-2-yl)carbamimidoyl)isonicotinimidamide(MTF-261)

Synthesized following the general procedure A using1-(4-(3-nitrophenyl)thiazol-2-yl)guanidine (1.00 g, 3.80 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 228 mg, 5.70 mmol) and4-pyridinecarbonitrile (395 mg, 3.80 mmol) to afford the titled compoundas a yellow powder (544 mg, 39%). ¹H NMR (500 MHz, DMSO-d6): δ 10.14(br. s, 1H), 9.17 (d, J=1.6 Hz, 1H), 8.81 (br. s, 2H), 8.73 (dd, J=4.7,1.4 Hz, 1H), 8.63 (s, 1H), 8.33 (t, J=8.4 Hz, 2H), 8.15 (dd, J=8.1, 1.5Hz, 1H), 7.81 (s, 1H), 7.76 (br. s, 1H), 7.72 (t, J=8.0 Hz, 1H), 7.54(dd, J=7.8, 4.8 Hz, 1H). ¹³C NMR (50 MHz, DMSO-d6): δ 173.30, 160.78,159.84, 151.81, 148.78, 148.32, 147.48, 136.02, 135.13, 131.79, 130.97,130.19, 123.25, 121.99, 119.79, 109.31. HRMS-ESI (m/z): [M+H]⁺ calc. forC₁₆H₁₃N₇O₂S⁺, 368.09242; Found: 368.09283. HPLC (λ₂₈₀): Purity 96.8%;t_(R): 6.367 min (method 1).

4-methoxy-N—(N-(4-phenylthiazol-2-yl)carbamimidoyl)benzimidamide(MTF-262)

Synthesized following the general procedure A using1-(4-phenylthiazol-2-yl)guanidine (1.00 g, 4.58 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 275 mg, 6.87 mmol) and4-methoxybenzonitrile (610 mg, 4.58 mmol) to afford the titled compoundas a beige powder (32 mg, 2%). ¹H NMR (400 MHz, DMSO-d6): δ 10.30 (br.s, 1H), 9.05 (br. s, 1H), 8.55 (s, 1H), 8.01 (d, J=8.9 Hz, 2H), 7.87 (d,J=7.3 Hz, 2H), 7.68 (br. s, 1H), 7.46 (s, 1H), 7.43 (t, J=7.7 Hz, 1H),7.31 (t, J=7.3 Hz, 1H), 7.04 (d, J=8.9 Hz, 2H), 3.83 (s, 3H). ¹³C NMR(101 MHz, DMSO-d6): δ 173.16, 161.77, 161.07, 160.84, 149.72, 134.54,129.21, 128.64, 127.50, 127.36, 125.51, 113.46, 106.11, 55.37, 38.89.HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₈H₁₇N₅OS⁺, 352.12266; Found:352.12268. HPLC (λ₂₈₀): Purity 95.1%; t_(R): 6.842 min (method 1).

N—(N-(4-phenylthiazol-2-yl)carbamimidoyl)picolinimidamide (MTF-263)

Synthesized following the general procedure A using1-(4-phenylthiazol-2-yl)guanidine (1.00 g, 4.58 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 275 mg, 6.87 mmol) and2-pyridinecarbonitrile (441 mL, 4.58 mmol) to afford the titled compoundas a yellow powder (369 mg, 25%). ¹H NMR (500 MHz, DMSO-d6): δ 10.16(br. s, 1H), 9.19 (br. s, 1H), 8.79 (br. s, 1H), 8.70 (d, J=3.6 Hz, 1H),8.38 (d, J=7.7 Hz, 1H), 8.01 (t, J=7.4 Hz, 1H), 8.19-7.70 (m, 4H), 7.88(d, br. s, J=7.4 Hz, 4H), 7.66-7.57 (m, 1H), 7.50 (s, 1H), 7.43 (t,J=7.3 Hz, 2H), 7.32 (t, J=7.0 Hz, 1H). ¹³C NMR (50 MHz, DMSO-d6): δ173.20, 160.91, 158.35, 150.73, 149.86, 148.61, 137.40, 134.50, 128.70,127.61, 126.38, 125.59, 122.04, 106.61. HRMS-ESI (m/z): [M+H]⁺ calc. forC₁₆H₁₄N₆S⁺, 323.10734; Found: 323.10770. HPLC (λ₂₈₀): Purity 98.0%;t_(R): 6.692 min (method 1).

3-bromo-N—(N-(6-methoxybenzo[d]thiazol-2-yl)carbamimidoyl)benzimidamide(MTF-264)

Synthesized following the general procedure A using1-(6-methoxybenzo[d]thiazol-2-yl)guanidine (1.00 g, 4.50 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 270 mg, 6.75 mmol) and3-bromobenzonitrile (819 mg, 4.50 mmol) to afford the titled compound asa dark gray powder (146 mg, 8%). ¹H NMR (500 MHz, DMSO-d6): δ 10.29 (br.s, 1H), 9.24 (br. s, 1H), 8.84 (br. s, 1H), 8.23 (s, 1H), 8.01 (d, br.s, J=7.5 Hz, 2H), 7.77 (d, J=7.2 Hz, 1H), 7.56 (d, J=8.6 Hz, 1H), 7.48(t, J=7.8 Hz, 1H), 7.42 (s, 1H), 6.95 (d, J=8.5 Hz, 1H), 3.79 (s, 3H).¹³C NMR (50 MHz, DMSO-d6): δ 170.29, 161.45, 160.30, 155.69, 145.55,137.47, 134.03, 132.39, 130.45, 130.36, 126.46, 121.68, 120.37, 113.92,104.89, 55.55. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₆H₁₄BrN₅OS⁺,404.01752; Found: 404.01733. HPLC (λ₂₈₀): Purity 95.2%; t_(R): 6.825 min(method 1).

3-bromo-N—(N-(6-methylbenzo[d]thiazol-2-yl)carbamimidoyl)benzimidamide(MTF-265)

Synthesized following the general procedure A using1-(6-methylbenzo[d]thiazol-2-yl)guanidine (1.00 g, 4.85 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 291 mg, 7.28 mmol) and3-bromobenzonitrile (883 mg, 4.85 mmol) to afford the titled compound asa beige powder (207 mg, 11%). ¹H NMR (500 MHz, DMSO-d6): δ 10.30 (br. s,1H), 9.32 (br. s, 1H), 8.87 (br. s, 1H), 8.24 (s, 1H), 8.01 (d, br. s,J=7.6 Hz, 2H), 7.77 (d, J=7.7 Hz, 1H), 7.60 (s, 1H), 7.54 (d, J=8.1 Hz,1H), 7.48 (t, J=7.8 Hz, 1H), 7.16 (d, J=8.0 Hz, 1H), 2.37 (s, 3H). ¹³CNMR (50 MHz, DMSO-d6): δ 171.42, 161.76, 160.43, 149.40, 137.44, 134.05,132.22, 131.30, 130.41, 126.86, 126.47, 121.69, 120.95, 119.45, 20.93.HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₆H₁₄BrN₅S⁺, 388.02261; Found:388.02313. HPLC (λ₂₈₀): Purity 98.3%; t_(R): 6.992 min (method 1).

3-chloro-N—(N-(4-phenylthiazol-2-yl)carbamimidoyl)benzimidamide(MTF-267)

Synthesized following the general procedure A using1-(4-phenylthiazol-2-yl)guanidine (1.00 g, 4.58 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 275 mg, 6.87 mmol) and3-chlorobenzonitrile (630 mg, 4.58 mmol) to afford the titled compoundas a white-yellowish powder (749 mg, 46%). ¹H NMR (500 MHz, DMSO-d6): δ10.36 (br. s, 1H), 9.15 (br. s, 1H), 8.78 (br. s, 1H), 8.10 (t, J=1.7Hz, 1H), 7.98 (d, J=7.9 Hz, 1H), 7.88 (d, br. s, J=7.2 Hz, 3H), 7.63(dd, J=8.0, 1.2 Hz, 1H), 7.54 (t, J=7.9 Hz, 1H), 7.48 (s, 1H), 7.43 (t,J=7.7 Hz, 2H), 7.31 (t, J=7.3 Hz, 1H). ¹³C NMR (50 MHz, DMSO-d6): δ173.05, 160.66, 160.04, 149.85, 137.43, 134.51, 133.20, 131.03, 130.14,128.69, 127.59, 127.46, 126.08, 125.58, 106.53. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₇H₁₄ClN₅S⁺, 356.07312; Found: 356.07321. HPLC (λ₂₈₀): Purity98.0%; t_(R): 7.008 min (method 1).

4-chloro-N—(N-(5-phenylthiazol-2-yl)carbamimidoyl)benzimidamide(MTF-268)

Synthesized following the general procedure A using1-(4-phenylthiazol-2-yl)guanidine (1.00 g, 4.58 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 275 mg, 6.87 mmol) and4-chlorobenzonitrile (630 mg, 4.58 mmol) to afford the titled compoundas a yellow powder (717 mg, 44%). ¹H NMR (500 MHz, DMSO-d6): δ 10.35(br. s, 1H), 9.14 (br. s, 1H), 8.73 (br. s, 1H), 8.05 (d, J=8.6 Hz, 2H),7.87 (d, J=7.2 Hz, 2H), 7.73 (br. s, 1H), 7.58 (d, J=8.6 Hz, 2H), 7.47(s, 1H), 7.43 (t, J=7.7 Hz, 2H), 7.31 (t, J=7.3 Hz, 1H). ¹³C NMR (50MHz, DMSO-d6): δ 173.07, 160.72, 160.43, 149.82, 136.13, 134.52, 134.17,129.37, 128.69, 128.31, 127.58, 125.57, 106.47. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₇H₁₄ClN₅S⁺, 356.07312; Found: 356.07318. HPLC (λ₂₈₀): Purity99.1%; t_(R): 6.925 min (method 1).

N—(N-(4-phenylthiazol-2-yl)carbamimidoyl)benzimidamide (MTF-272)

Synthesized following the general procedure A using1-(4-phenylthiazol-2-yl)guanidine (1.00 g, 4.58 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 275 mg, 6.87 mmol) andbenzonitrile (0.471 mL, 4.58 mmol) to afford the titled compound as abeige powder (486 mg, 33%). ¹H NMR (500 MHz, DMSO-d6): δ 10.33 (br. s,1H), 9.13 (br. s, 1H), 8.67 (br. s, 1H), 8.02 (d, J=7.2 Hz, 2H), 7.88(d, J=7.3 Hz, 2H), 7.72 (br. s, 1H) 7.56 (t, J=7.2 Hz, 1H), 7.49 (dd,J=14.9, 7.7 Hz, 3H), 7.43 (t, J=7.7 Hz, 2H), 7.31 (t, J=7.3 Hz, 1H). ¹³CNMR (50 MHz, DMSO-d6): δ 173.16, 161.66, 160.90, 149.80, 135.41, 134.54,131.26, 128.71 (2C), 128.22 (2C), 127.57, 127.50 (2C), 125.57 (2C),106.37. HPLC (λ₂₈₀): Purity 82.0%; t_(R): 6.850 min (method 1).

N—(N-(4-phenylthiazol-2-yl)carbamimidoyl)nicotinimidamide (MTF-273)

Synthesized following the general procedure A using1-(4-phenylthiazol-2-yl)guanidine (1.00 g, 4.58 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 275 mg, 6.87 mmol) and3-cyanopyridine (477 mg, 4.58 mmol) to afford the titled compound as ayellowish powder (694 mg, 47%). ¹H NMR (500 MHz, DMSO-d6): δ 10.30 (br.s, 1H), 9.17 (d, br. s, J=1.6 Hz, 2H), 8.82 (br. s, 1H), 8.73 (dd,J=4.7, 1.4 Hz, 1H), 8.33 (d, J=8.0 Hz, 1H), 7.88 (d, br. s, J=7.3 Hz,3H), 7.54 (dd, J=7.8, 4.8 Hz, 1H), 7.49 (s, 1H), 7.43 (t, J=7.6 Hz, 2H),7.32 (t, J=7.3 Hz, 1H). ¹³C NMR (50 MHz, DMSO-d6): δ 173.07, 160.68,159.88, 151.80, 149.86, 148.77, 135.10, 134.52, 131.05, 128.71, 127.60,125.59, 123.25, 106.58. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₆H₁₄N₆S⁺,323.10734; Found: 323.10764. HPLC (λ₂₈₀): Purity 96.9%; t_(R): 6.075 min(method 1).

N—(N-(4-phenylthiazol-2-yl)carbamimidoyl)isonicotinimidamide (MTF-274)

Synthesized following the general procedure A using1-(4-phenylthiazol-2-yl)guanidine (1.00 g, 4.58 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 275 mg, 6.87 mmol) and4-pyridinecarbonitrile (477 mg, 4.58 mmol) to afford the titled compoundas a yellowish powder (871 mg, 59%). ¹H NMR (500 MHz, DMSO-d6): δ 10.30(br. s, 1H), 9.20 (br. s, 1H), 8.86 (br. s, 1H), 8.75 (dd, J=4.5, 1.6Hz, 2H), 7.92 (dd, J=4.5, 1.6 Hz, 2H), 7.88 (d, br. s, J=7.2 Hz, 3H),7.50 (s, 1H), 7.43 (t, J=7.7 Hz, 2H), 7.32 (t, J=7.3 Hz, 1H). ¹³C NMR(50 MHz, DMSO-d6): δ 173.00, 160.64, 159.63, 150.09, 149.90, 142.74,134.50, 128.72, 127.63, 125.61, 121.49, 106.75. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₆H₁₄N₆S⁺, 323.10734; Found: 323.10757. HPLC (λ₂₈₀): Purity95.2%; t_(R): 6.058 min (method 1).

2-chloro-N—(N-(4-(3-nitrophenyl)thiazol-2-yl)carbamimidoyl)benzimidamide(MTF-276)

Synthesized following the general procedure A using1-(4-(3-nitrophenyl)thiazol-2-yl)guanidine (1.00 g, 3.80 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 228 mg, 5.70 mmol) and2-chlorobenzonitrile (523 mg, 3.80 mmol) to afford the titled compoundas a brown powder (30 mg, 2%). ¹H NMR (400 MHz, DMSO-d6): δ 10.08 (br.s, 1H), 8.83 (br. s, 1H), 8.70 (br. s, 1H), 8.62 (s, 1H), 8.34 (d, J=7.7Hz, 1H), 8.15 (d, J=7.4 Hz, 1H), 8.03-7.64 (br. s, 1H), 7.81 (s, 1H),7.72 (t, J=7.7 Hz, 1H), 7.52 (t, J=7.8 Hz, 2H), 7.49-7.37 (m, 2H). ¹³CNMR (101 MHz, DMSO-d6): δ 173.55, 162.25, 160.72, 148.36, 147.36,136.69, 136.00, 131.76, 130.52, 130.25 (2C), 129.51, 129.39, 126.87,121.95, 119.70, 109.25. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₇H₁₃ClN₆O₂S⁺,401.05820; Found: 401.05820. HPLC (λ₂₈₀): Purity 95.2%; t_(R): 6.858 min(method 1).

3-chloro-N—(N-(4-(3-nitrophenyl)thiazol-2-yl)carbamimidoyl)benzimidamide(MTF-277)

Synthesized following the general procedure A using1-(4-(3-nitrophenyl)thiazol-2-yl)guanidine (1.00 g, 3.80 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 228 mg, 5.70 mmol) and3-chlorobenzonitrile (523 mg, 3.80 mmol) to afford the titled compoundas a brown powder (457 mg, 30%). ¹H NMR (500 MHz, DMSO-d6): δ 10.22 (br.s, 1H), 8.77 (br. s, 1H), 8.62 (s, 1H), 8.34 (d, J=7.7 Hz, 1H),8.23-7.91 (br. s, 1H), 8.15 (d, J=8.0 Hz, 1H), 8.09 (s, 1H), 7.97 (d,J=7.7 Hz, 1H), 7.80 (s, 1H), 7.72 (t, J=8.0 Hz, 1H), 7.63 (d, J=7.9 Hz,1H), 7.54 (t, J=7.9 Hz, 1H). ¹³C NMR (50 MHz, DMSO-d6): δ 173.31,160.75, 160.02, 148.34, 147.48, 137.34, 136.02, 133.17, 131.83, 131.05,130.24, 130.17, 127.45, 126.11, 122.02, 119.78, 109.32. HRMS-ESI (m/z):[M+H]⁺ calc. for C₁₇H₁₃ClN₆O₂S⁺, 401.05820; Found: 401.05835. HPLC(λ₂₈₀): Purity 95.5%; t_(R): 7.025 min (method 1).

4-chloro-N—(N-(4-(3-nitrophenyl)thiazol-2-yl)carbamimidoyl)benzimidamide(MTF-281)

Synthesized following the general procedure A using1-(4-(3-nitrophenyl)thiazol-2-yl)guanidine (1.00 g, 3.80 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 228 mg, 5.70 mmol) and4-chlorobenzonitrile (523 mg, 3.80 mmol) to afford the titled compoundas a yellow powder (640 mg, 42%). ¹H NMR (200 MHz, DMSO-d6): δ 10.15(br. s, 1H), 8.74 (br. s, 2H), 8.68-8.60 (m, 1H), 8.55-7.90 (br. s, 1H),8.39-8.30 (m, 1H), 8.16 (ddd, J=8.2, 2.3, 0.8 Hz, 1H), 8.10-7.97 (m,2H), 7.82 (s, 1H), 7.72 (t, J=8.0 Hz, 1H), 7.66-7.48 (m, 2H). ¹³C NMR(50 MHz, DMSO-d6): δ 173.37, 160.84, 160.46, 148.30, 147.47, 136.17,136.04, 134.10, 131.77, 130.16, 129.40 (2C), 128.29 (2C), 121.96,119.77, 109.19. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₇H₁₃ClN₆O₂S⁺,401.05820; Found: 401.05820. HPLC (λ₂₈₀): Purity 96.7%; t_(R): 7.033 min(method 1).

N—(N-(4-methylthiazol-2-yl)carbamimidoyl)benzimidamide (MTF-283)

Synthesized following the general procedure A using1-(4-methylthiazol-2-yl)guanidine (1.00 g, 6.41 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 384 mg, 9.61 mmol) andbenzonitrile (0.66 mL, 6.41 mmol) to afford the titled compound as abrown powder (382 mg, 23%). ¹H NMR (500 MHz, DMSO-d6): δ 10.42 (br. s,1H), 9.11 (br. s, 1H), 8.61 (br. s, 1H), 8.02-7.96 (m, 2H), 7.57-7.52(m, 1H), 7.85-7.39 (br. s, 4H), 7.51-7.46 (m, 2H), 6.57 (d, J=1.0 Hz,1H), 2.24 (d, J=0.9 Hz, 3H). ¹³C NMR (50 MHz, DMSO-d6): δ 172.99,161.63, 160.69, 147.64, 135.50, 131.20, 128.21, 127.45, 105.64, 17.56.HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₂H₁₃N₅S⁺, 260.09644; Found:260.09653. HPLC (λ₂₈₀): Purity 95.7%; t_(R): 5.167 min (method 1).

2-chloro-N—(N-(4-methylthiazol-2-yl)carbamimidoyl)benzimidamide(MTF-284)

Synthesized following the general procedure A using1-(4-methylthiazol-2-yl)guanidine (1.00 g, 6.41 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 384 mg, 9.61 mmol) and2-chlorobenzonitrile (881 mg, 6.41 mmol) to afford the titled compoundas a white powder (132 mg, 7%). ¹H NMR (400 MHz, DMSO-d6): δ 10.27 (br.s, 1H), 9.06 (br. s, 1H), 8.63 (br. s, 1H), 7.63 (br. s, 1H), 7.46 (m,4H), 6.57 (s, 1H), 2.24 (s, 3H). ¹³C NMR (101 MHz, DMSO-d6): δ 172.87,162.18, 160.47, 147.49, 136.79, 130.49, 130.27, 129.55, 129.40, 126.88,105.69, 17.46. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₂H₁₂ClN₅S⁺, 294.05747;Found: 294.05747. HPLC (λ₂₈₀): Purity 95.0%; t_(R): 5.500 min (method1).

3-chloro-N—(N-(4-methylthiazol-2-yl)carbamimidoyl)benzimidamide(MTF-285)

Synthesized following the general procedure A using1-(4-methylthiazol-2-yl)guanidine (1.00 g, 6.41 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 384 mg, 9.61 mmol) and3-chlorobenzonitrile (881 mg, 6.41 mmol) to afford the titled compoundas a white-yellowish (1.24 g, 66%). ¹H NMR (500 MHz, DMSO-d6): δ 10.43(br. s, 1H), 9.13 (br. s, 1H), 8.71 (br. s, 1H), 8.11-8.05 (m, 1H), 7.95(d, J=7.9 Hz, 1H), 7.64-7.59 (m, 1H), 7.58 (br.s, 1H), 7.53 (t, J=7.9Hz, 1H), 6.59 (d, J=0.9 Hz, 1H), 2.25 (d, J=0.5 Hz, 3H). ¹³C NMR (50MHz, DMSO-d6): δ 172.82, 160.44, 159.91, 147.64, 137.47, 133.18, 130.97,130.15, 127.39, 126.00, 105.82, 17.53. HRMS-ESI (m/z): [M+H]⁺ calc. forC₁₂H₁₂ClN₅S⁺, 294.05747; Found: 294.05768. HPLC (λ₂₈₀): Purity 95.0%;t_(R): 5.933 min (method 1).

4-chloro-N—(N-(4-methylthiazol-2-yl)carbamimidoyl)benzimidamide(MTF-286)

Synthesized following the general procedure A using1-(4-methylthiazol-2-yl)guanidine (1.00 g, 6.41 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 384 mg, 9.61 mmol) and4-chlorobenzonitrile (881 mg, 6.41 mmol) to afford the titled compoundas a white-yellowish (923 mg, 49%). ¹H NMR (500 MHz, DMSO-d6): δ 10.42(br. s, 1H), 9.13 (br. s, 1H), 8.67 (br. s, 1H), 8.01 (d, J=8.6 Hz, 2H),7.57 (d, br. s J=8.6 Hz, 3H), 6.57 (d, J=1.0 Hz, 1H), 2.24 (d, J=0.7 Hz,3H). ¹³C NMR (50 MHz, DMSO-d6): δ 172.87, 160.50, 160.37, 147.63,136.08, 134.23, 129.30, 128.29, 105.74, 17.53. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₂H₁₂ClN₅S⁺, 294.05747; Found: 294.05762. HPLC (λ₂₈₀): Purity95.3%; t_(R): 5.958 min (method 1).

2-bromo-N—(N-(4-methylthiazol-2-yl)carbamimidoyl)benzimidamide (MTF-287)

Synthesized following the general procedure A using1-(4-methylthiazol-2-yl)guanidine (1.00 g, 6.41 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 384 mg, 9.61 mmol) and2-bromobenzonitrile (1.17 g, 6.41 mmol) to afford the titled compound asa white-yellowish (499 mg, 23%). ¹H NMR (400 MHz, DMSO-d6): δ 10.27 (br.s, 1H), 9.07 (br. s, 1H), 8.63 (br. s, 1H), 7.67 (d, br. s, J=7.9 Hz,2H), 7.46-7.43 (m, 2H), 7.36 (ddd, J=8.1, 6.0, 3.2 Hz, 1H), 6.57 (s,1H), 2.24 (s, 3H). ¹³C NMR (101 MHz, DMSO-d6): δ 172.89, 163.29, 160.49,147.50, 138.85, 132.49, 130.57, 129.44, 127.37, 119.62, 105.68, 17.46.HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₂H₁₂BrN₅S⁺, 338.00696; Found:338.00705. HPLC (λ₂₈₀): Purity 95.8%; t_(R): 5.192 min (method 1).

3-bromo-N—(N-(4-methylthiazol-2-yl)carbamimidoyl)benzimidamide (MTF-288)

Synthesized following the general procedure A using1-(4-methylthiazol-2-yl)guanidine (1.00 g, 6.41 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 384 mg, 9.61 mmol) and3-bromobenzonitrile (1.17 g, 6.41 mmol) to afford the titled compound asa white-yellowish (1.11 g, 51%). ¹H NMR (500 MHz, DMSO-d6): δ 10.42 (s,1H), 9.12 (s, 1H), 8.72 (s, 1H), 8.21 (t, J=1.7 Hz, 1H), 7.98 (d, J=8.0Hz, 1H), 7.75 (dd, br. s, J=8.0, 1.1 Hz, 2H), 7.46 (t, J=7.9 Hz, 1H),6.59 (d, J=0.9 Hz, 1H), 2.24 (d, J=0.8 Hz, 3H). ¹³C NMR (50 MHz,DMSO-d6): δ 172.80, 160.42, 159.83, 147.63, 137.63, 133.86, 130.38,130.27, 126.35, 121.66, 105.81, 17.53. HRMS-ESI (m/z): [M+H]⁺ calc. forC₁₂H₁₂BrN₅S⁺, 338.00696; Found: 338.00760. HPLC (λ₂₈₀): Purity 95.1%;t_(R): 6.042 min (method 1).

N—(N-(4-methylthiazol-2-yl)carbamimidoyl)picolinimidamide (MTF-289)

Synthesized following the general procedure A using1-(4-methylthiazol-2-yl)guanidine (1.00 g, 6.41 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 384 mg, 9.61 mmol) and2-pyridinecarbonitrile (0.617 mL, 6.41 mmol) to afford the titledcompound as a yellow powder (1.05 g, 63%). ¹H NMR (200 MHz, DMSO-d6): δ10.20 (s, 1H), 9.15 (s, 1H), 8.77 (s, 1H), 8.73-8.65 (m, 1H), 8.34 (d,J=8.5 Hz, 1H), 8.00 (td, J=7.8, 1.7 Hz, 1H), 7.61 (ddd, J=7.4, 4.8, 1.1Hz, 2H), 6.60 (d, J=1.0 Hz, 1H), 2.25 (s, 3H). ¹³C NMR (101 MHz,DMSO-d6): δ 172.90, 160.61, 158.17, 150.75, 148.52, 147.60, 137.32,126.26, 121.87, 105.82, 17.46. HRMS-ESI (m/z): [M+H]⁺ calc. forC₁₁H₁₂N₆S⁺, 261.09169; Found: 261.09174. HPLC (λ₂₈₀): Purity 95.7%;t_(R): 5.708 min (method 1).

N—(N-(4-methylthiazol-2-yl)carbamimidoyl)nicotinimidamide (MTF-290)

Synthesized following the general procedure A using1-(4-methylthiazol-2-yl)guanidine (1.00 g, 6.41 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 384 mg, 9.61 mmol) and3-pyridinecarbonitrile (0.667 mg, 6.41 mmol) to afford the titledcompound as a yellow powder (751 mg, 45%). ¹H NMR (500 MHz, DMSO-d6): δ10.39 (br. s, 1H), 9.15 (d, br. s, J=1.7 Hz, 2H), 8.77 (br. s, 1H), 8.71(dd, J=4.7, 1.5 Hz, 1H), 8.36-8.23 (m, 1H), 7.65 (br. s, 1H), 7.52 (dd,J=7.5, 4.8 Hz, 1H), 6.59 (d, J=0.7 Hz, 1H), 2.25 (s, 3H). ¹³C NMR (50MHz, DMSO-d6): δ 172.84, 160.46, 159.79, 151.75, 148.72, 147.67, 135.03,131.09, 123.24, 105.89, 17.54. HRMS-ESI (m/z): [M+H]⁺ calc. forC₁₁H₁₂N₆S⁺, 261.09169; Found: 261.09171. HPLC (λ₂₈₀): Purity 97.1%;t_(R): 5.508 min (method 1).

N—(N-(4-methylthiazol-2-yl)carbamimidoyl)isonicotinimidamide (MTF-291)

Synthesized following the general procedure A using1-(4-methylthiazol-2-yl)guanidine (1.00 g, 6.41 mmol), sodium hydride(60% dispersion in mineral oil, 384 mg, 9.61 mmol) and4-pyridinecarbonitrile (0.667 mg, 6.41 mmol) to afford the titledcompound as a yellow powder (985 mg, 59%). ¹H NMR (500 MHz, DMSO-d6): δ10.37 (br. s, 1H), 9.15 (br. s, 1H), 8.80 (br. s, 1H), 8.74 (dd, J=4.5,1.6 Hz, 2H), 7.89 (dd, J=4.5, 1.6 Hz, 2H), 7.68 (br. s, 1H), 6.60 (d,J=0.9 Hz, 1H), 2.25 (d, J=0.8 Hz, 3H). ¹³C NMR (50 MHz, DMSO-d6): δ172.77, 160.43, 159.58, 150.08, 147.74, 142.81, 121.45, 106.08, 17.54.HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₁H₁₂N₆S⁺, 261.09169; Found:261.09174. HPLC (λ₂₈₀): Purity 99.5%; t_(R): 4.525 min (method 1).

4-methoxy-N—(N-(4-methylthiazol-2-yl)carbamimidoyl)benzimidamide(MTF-292)

Synthesized following the general procedure A using1-(4-methylthiazol-2-yl)guanidine (1.00 g, 6.41 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 384 mg, 9.61 mmol) and4-methoxybenzonitrile (0.853 mg, 6.41 mmol) to afford the titledcompound as a beige powder (167 mg, 9%). ¹H NMR (500 MHz, DMSO-d6): δ10.40 (br. s, 1H), 9.06 (br. s, 1H), 8.50 (br. s, 1H), 7.98 (d, J=8.9Hz, 2H), 7.49 (br. s, 1H), 7.02 (d, J=8.9 Hz, 2H), 6.55 (d, J=0.8 Hz,1H), 3.83 (s, 3H), 2.24 (s, 3H). ¹³C NMR (101 MHz, DMSO-d6): δ 172.97,161.71, 160.99, 160.62, 147.51, 129.13, 127.43, 113.43, 105.33, 55.34,17.48. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₃H₁₅N₅OS⁺, 290.10701; Found:290.10709. HPLC (λ₂₈₀): Purity 100.0%; t_(R): 5.308 min (method 1).

2-bromo-N—(N-(4,5-dimethylthiazol-2-yl)carbamimidoyl)benzimidamide(MTF-295)

Synthesized following the general procedure A using1-(4,5-dimethylthiazol-2-yl)guanidine (1.00 g, 5.88 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 353 mg, 8.82 mmol) and2-bromobenzonitrile (1.07 g, 5.88 mmol) to afford the titled compound asa brown powder (746 mg, 36%). ¹H NMR (500 MHz, DMSO-d6): δ 10.28 (br. s,1H), 9.01 (br. s, 1H), 8.58 (br. s, 1H), 7.70-7.30 (br. s, 1H), 7.67 (d,J=7.9 Hz, 1H), 7.44 (dd, J=8.1, 5.2 Hz, 2H), 7.40-7.33 (m, 1H), 2.21 (s,3H), 2.14 (s, 3H). ¹³C NMR (101 MHz, DMSO-d6): δ 169.26, 163.14, 160.22,142.62, 138.85, 132.48, 130.54, 129.44, 127.36, 119.64, 116.98, 14.58,10.71. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₃H₁₄BrN₅S⁺, 352.02261; Found:352.02289. HPLC (λ₂₈₀): Purity 97.8%; t_(R): 5.717 min (method 1).

2-bromo-N—(N-(4,5-dimethylthiazol-2-yl)carbamimidoyl)benzimidamide(MTF-296)

Synthesized following the general procedure A using1-(4,5-dimethylthiazol-2-yl)guanidine (1.00 g, 5.88 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 353 mg, 8.82 mmol) and3-bromobenzonitrile (1.07 g, 5.88 mmol) to afford the titled compound asa brown powder brown powder (746 mg, 36%). ¹H NMR (200 MHz, DMSO-d6): δ10.45 (br. s, 1H), 9.01 (br. s, 1H), 8.70 (br. s, 1H), 8.20 (t, J=1.7Hz, 1H), 8.06-7.28 (br. s, 1H), 7.97 (d, J=8.0 Hz, 1H), 7.74 (ddd,J=7.9, 1.9, 0.8 Hz, 1H), 7.46 (t, J=7.9 Hz, 1H), 2.21 (s, 3H), 2.15 (s,3H). ¹³C NMR (50 MHz, DMSO-d6): δ 169.14, 160.12, 159.63, 142.74,137.65, 133.81, 130.41, 130.18, 126.29, 121.61, 117.13, 14.63, 10.76.HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₃H₁₅BrN₅S⁺, 352.02261; Found:352.02377. HPLC (λ₂₈₀): Purity 97.3%; t_(R): 5.767 min (method 1).

N—(N-(4,5-dimethylthiazol-2-yl)carbamimidoyl)picolinimidamide (MTF-297)

Synthesized following the general procedure A using1-(4,5-dimethylthiazol-2-yl)guanidine (1.00 g, 5.88 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 353 mg, 8.82 mmol) and2-pyridinecarbonitrile (0.566 mL, 5.88 mmol) to afford the titledcompound as a yellow powder (726 mg, 45%). ¹H NMR (500 MHz, DMSO-d6): δ10.19 (br. s, 1H), 9.09 (br. s, 1H), 8.68 (d, br. s. J=4.1 Hz, 2H), 8.33(d, J=7.9 Hz, 1H), 7.99 (td, J=7.7, 1.3 Hz, 1H), 7.60 (dd, br. s, J=6.5,5.0 Hz, 2H), 2.21 (s, 3H), 2.15 (s, 3H). ¹³C NMR (101 MHz, DMSO-d6): δ169.29, 160.32, 158.01, 150.80, 148.50, 142.71, 137.28, 126.19, 121.80,117.16, 14.56, 10.69. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₂H₁₄N₆S⁺,275.10734; Found: 275.10742. HPLC (λ₂₈₀): Purity 100.0%; t_(R): 5.475min (method 1).

N—(N-(4,5-dimethylthiazol-2-yl)carbamimidoyl)nicotinimidamide (MTF-298)

Synthesized following the general procedure A using1-(4,5-dimethylthiazol-2-yl)guanidine (1.00 g, 5.88 mmol), sodiumhydride (60% dispersion in mineral oil, 353 mg, 8.82 mmol) and3-pyridinecarbonitrile (612 mg, 5.88 mmol) to afford the titled compoundas a yellow powder (742 mg, 46%). ¹H NMR (500 MHz, DMSO-d6): δ 10.43(br. s, 1H), 9.04 (br. s, 1H), 8.67 (br. s, 1H), 8.20 (t, J=1.6 Hz, 1H),7.97 (d, J=8.0 Hz, 1H), 7.80-7.70 (m, 1H), 7.88-7.29 (br. s, 1H), 7.46(t, J=7.9 Hz, 1H), 2.21 (s, 3H), 2.15 (s, 3H). ¹³C NMR (101 MHz,DMSO-d6): δ 169.13, 160.11, 159.55, 151.63, 148.60, 142.72, 134.89,131.06, 123.17, 117.16, 14.57, 10.69. HRMS-ESI (m/z): [M+H]⁺ calc. forC₁₂H₁₄N₆S⁺, 275.10734; Found: 275.10736. HPLC (λ₂₈₀): Purity 97.0%;t_(R): 5.458 min (method 1).

N—(N-(4,5-dimethylthiazol-2-yl)carbamimidoyl)isonicotinimidamide(MTF-299)

Synthesized following the general procedure A using1-(4,5-dimethylthiazol-2-yl)guanidine (1.00 g, 5.88 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 353 mg, 8.82 mmol) and4-pyridinecarbonitrile (612 mg, 5.88 mmol) to afford the titled compoundas a yellow powder (548 mg, 34%). ¹H NMR (500 MHz, DMSO-d6): δ 10.40(br. s, 1H), 9.09 (br. s, 1H), 8.73 (d, br. s., J=5.7 Hz, 3H), 7.88 (d,J=5.5 Hz, 2H), 7.61 (br. s, 1H), 2.21 (s, 3H), 2.15 (s, 3H). ¹³C NMR(101 MHz, DMSO-d6): δ 169.05, 160.06, 159.33, 149.98, 142.78, 121.31,117.38, 14.55, 10.68. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₂H₁₄N₆S⁺,275.10734; Found: 275.10739. HPLC (λ₂₈₀): Purity 99.5%; t_(R): 5.108 min(method 1).

2-chloro-N—(N-(4,5-dimethylthiazol-2-yl)carbamimidoyl)benzimidamide(MTF-300)

Synthesized following the general procedure A using1-(4,5-dimethylthiazol-2-yl)guanidine (1.00 g, 5.88 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 353 mg, 8.82 mmol) and2-chlorobenzonitrile (809 mg, 5.88 mmol) to afford the titled compoundas a yellow powder (109 mg, 6%). ¹H NMR (500 MHz, DMSO-d6): δ 10.30 (br.s, 1H), 9.01 (br. s, 1H), 8.59 (br. s, 1H), 7.46 (m, 5H), 2.21 (s, 3H),2.14 (s, 3H). ¹³C NMR (101 MHz, DMSO-d6): δ 169.26, 162.01, 160.22,142.63, 136.82, 130.47, 130.28, 129.56, 129.39, 126.88, 117.00, 14.58,10.71. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₃H₁₄CN₅S⁺, 308.07312; Found:308.07318. HPLC (λ₂₈₀): Purity 95.4%; t_(R): 5.425 min (method 1).

3-chloro-N—(N-(4,5-dimethylthiazol-2-yl)carbamimidoyl)benzimidamide(MTF-301)

Synthesized following the general procedure A using1-(4,5-dimethylthiazol-2-yl)guanidine (1.00 g, 5.88 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 353 mg, 8.82 mmol) and3-chlorobenzonitrile (809 mg, 5.88 mmol) to afford the titled compoundas a yellow powder (416 mg, 23%). ¹H NMR (500 MHz, DMSO-d6): δ 10.44(br. s, 1H), 9.04 (br. s, 1H), 8.68 (br. s, 1H), 8.08-8.03 (m, 1H), 7.93(d, J=7.9 Hz, 1H), 7.61 (ddd, J=8.0, 2.1, 0.9 Hz, 1H), 7.78-7.39 (br. s,1H), 7.52 (t, J=7.9 Hz, 1H), 2.21 (s, 3H), 2.15 (s, 3H). ¹³C NMR (50MHz, DMSO-d6): δ 169.15, 160.14, 159.71, 142.73, 137.49, 133.13, 130.91,130.15, 127.31, 125.93, 117.14, 14.62, 10.75. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₃H₁₄CN₅S⁺, 308.07312; Found: 308.07321. HPLC (λ₂₈₀): Purity95.2%; t_(R): 6.075 min (method 1).

4-chloro-N—(N-(4,5-dimethylthiazol-2-yl)carbamimidoyl)benzimidamide(MTF-302)

Synthesized following the general procedure A using1-(4,5-dimethylthiazol-2-yl)guanidine (1.00 g, 5.88 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 353 mg, 8.82 mmol)4-chlorobenzonitrile (809 mg, 5.88 mmol to afford the titled compound asa yellow powder (90 mg, 5%). ¹H NMR (500 MHz, DMSO-d6): δ 10.44 (br. s,1H), 9.04 (br. s, 1H), 8.64 (br. s, 1H), 8.01 (d, J=8.6 Hz, 2H), 7.57(d, br. s, J=8.6 Hz, 3H), 2.22 (s, 3H), 2.15 (s, 3H). ¹³C NMR (101 MHz,DMSO-d6): δ 169.15, 160.16, 160.11, 142.66, 135.93, 134.23, 129.19 (2C),128.22 (2C), 117.02, 14.57, 10.70. HRMS-ESI (m/z): [M+H]⁺ calc. forC₁₃H₁₄ClN₅S⁺, 308.07312; Found: 308.07315. HPLC (λ₂₈₀): Purity 98.0%;t_(R): 6.267 min (method 1).

N—(N-(4,5-dimethylthiazol-2-yl)carbamimidoyl)benzimidamide (MTF-303)

Synthesized following the general procedure A using1-(4,5-dimethylthiazol-2-yl)guanidine (1.00 g, 5.88 mmol), sodiumhydride (60% dispersion in mineral oil, 1.5 eq., 353 mg, 8.82 mmol) andbenzonitrile (0.605 mL, 5.88 mmol) to afford the titled compound as ayellow powder (113 mg, 7%). ¹H NMR (200 MHz, DMSO-d6): δ 10.45 (s, 1H),8.94 (s, 1H), 8.61 (s, 1H), 7.98 (dd, J=7.9, 1.7 Hz, 2H), 7.84-7.31 (m,4H), 2.21 (d, J=0.6 Hz, 3H), 2.15 (s, 3H). ¹³C NMR (101 MHz, DMSO-d6): δ169.26, 161.31, 160.33, 142.65, 135.47, 131.06, 128.12, 127.30, 116.88,14.58, 10.70. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₃H₁₅N₅S⁺, 274.11209;Found: 274.11212. HPLC (λ₂₈₀): Purity 97.5%; t_(R): 5.408 min (method1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2,2,2-trichloroacetimidamide(MTF-305)

A solution of 1-(benzo[d]thiazol-2-yl)guanidine (500 mg, 2.6 mmol) andtrichloroacetonitrile (260 μL, 2.6 mmol) in technical grade ethanol (5mL) was stirred at r.t. under argon atmosphere for 22 h. The precipitatethat formed was filtered, washed with little amount of cold ethanol,then with a very little amount of diethyl ether, dried quickly at air(the compound oxidized over time at air) and stored under argon. Yellowsolid (100 mg, 11%). TLC: R_(f) (CHCl₃/MeOH, 95/5, v/v)=0.90. ¹H NMR(400 MHz, Acetone-d6): δ 10.71 (s, 1H), 9.84 (s, 1H), 8.39 (s, 1H), 7.80(dd, J=7.9, 1.3 Hz, 1H), 7.72 (d, J=8.0 Hz, 1H), 7.66 (s, 1H), 7.38 (td,J=8.2, 7.8, 1.3 Hz, 1H), 7.25 (td, J=7.6, 1.2 Hz, 1H). ¹³C NMR (101 MHz,Acetone-d6): δ 206.12, 126.63, 124.28, 121.93, 121.33. HRMS-ESI (m/z):[M+H]⁺ calc. for C₁₀H₉Cl₃N₅S⁺, 335.96388; Found: 335.96420. HPLC (λ₂₅₄):Purity 99.7%; t_(R): 7.983 min (method 1).

N2-(2-(methylthio)phenyl)-6-(trichloromethyl)-1,3,5-triazine-2,4-diamine(MTF-316)

To suspension ofN2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(trichloromethyl)-1,3,5-triazine-2,4-diamine)CRO15 (150 mg, 0.225 mmol) in methanol (10 mL) was added mercaptoethanol(350 μL, 4.5 mmol) and the mixture stirred at r.t. After 10 minstirring, total solubility was reached. TLC and LCMS showed totalconversion into the reduced thiophenol. After 20 h stirring at r.t.,K₂CO₃ (62.1 mg, 0.45 mmol) was added and the mixture sonicated untiltotal solubilisation. Then MeI (28 μL, 0.45 mmol) was added and thesolution stirred for 5 h at r.t. (reaction time not optimized). TLC andLCMS showed total conversion into two new compounds (ratio 2/1). Themixture was extracted with EtOAc twice and the combined organic layerswere dried with Na₂SO₄ and evaporated under reduced pressure.Purification by silicagel flash chromatography (cyclohexane/EtOAc, 10/0to 7/3, v/v) afforded the desired compound as white crystals (27.0 mg,34% over 2 steps). ¹H NMR (200 MHz, Acetone-d6): S 8.04 (s, 1H), 7.73(d, J=7.9 Hz, 1H), 7.11 (dd, J=7.7, 1.7 Hz, 1H), 7.01-6.57 (m, 4H), 2.05(s, 3H). ¹³C NMR (50 MHz, Acetone-d6): δ 174.04, 168.91, 166.25, 138.04,131.38, 130.32, 128.05, 125.84, 124.21, 97.43. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₁H₁₁Cl₃N₅S⁺, 349.97953; Found: 349.97983. HPLC (λ₂₅₄):Purity 97.6%; t_(R): 10.483 min (method 2).

6-(dichloromethyl)-N2-(2-(methylthio)phenyl)-1,3,5-triazine-2,4-diamine(MTF-317)

To suspension ofN2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(trichloromethyl)-1,3,5-triazine-2,4-diamine)CRO15 (150 mg, 0.225 mmol) in methanol (10 mL) was added mercaptoethanol(350 μL, 4.5 mmol) and the mixture stirred at r.t. After 10 minstirring, total solubility was reached. TLC and LCMS showed totalconversion into the reduced thiophenol. After 20 h stirring at r.t.,K₂CO₃ (62.1 mg, 0.45 mmol) was added and the mixture sonicated untiltotal solubilisation. Then MeI (28 μL, 0.45 mmol) was added and thesolution stirred for 5 h at r.t. (reaction time not optimized). TLC andLCMS showed total conversion into two new compounds (ratio 2/1). Themixture was extracted with EtOAc twice and the combined organic layerswere dried with Na₂SO₄ and evaporated under reduced pressure.Purification by silicagel flash chromatography (cyclohexane/EtOAc, 10/0to 7/3, v/v) afforded the titled compound as white crystals (12.7 mg,18% over 2 steps). ¹H NMR (200 MHz, Acetone-d6): δ 8.27 (s, 1H),8.19-8.07 (m, 1H), 7.49 (dd, J=7.6, 1.7 Hz, 1H), 7.32-7.23 (m, 1H), 7.16(td, J=7.5, 1.5 Hz, 1H), 6.90 (s, 2H), 6.54 (s, 1H), 2.43 (s, 3H). ¹³CNMR (50 MHz, Acetone-d6): δ 173.85, 168.61, 166.05, 138.23, 131.54,130.28, 128.11, 125.67, 124.06, 71.59, 17.73. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₁H₁₂Cl₂N₅S⁺, 316.01850; Found: 316.01892. HPLC (λ₂₅₄):Purity 97.3%; t_(R): 10.658 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(3-methoxyphenyl)-1,3,5-triazine-2,4-diamine(MTF-318)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-methoxybenzimidamide (325mg, 1 mmol) to afford the titled compound as a white powder (272 mg,84%). ¹H NMR (200 MHz, DMSO-d6): δ 9.25 (s, 2H), 7.82 (dd, J=8.1, 5.1Hz, 4H), 7.61 (dd, J=7.7, 1.4 Hz, 2H), 7.44-7.33 (m, 4H), 7.27 (td,J=7.5, 1.4 Hz, 2H), 7.22-6.97 (m, 8H), 3.78 (s, 6H). ¹³C NMR (50 MHz,DMSO-d6): δ 169.95, 167.28, 165.48, 159.19, 138.15, 136.69, 133.74,129.31, 128.36, 127.63, 126.62, 126.32, 120.25, 117.26, 112.78, 55.10.HRMS-ESI (m/z): [M+H]+ calc. for C₁₅H₁₂BrN₅S⁺, 374.00696; Found:374.00797. HPLC (km): Purity 100.0%; t_(R): 11.458 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(3-ethoxyphenyl)-1,3,5-triazine-2,4-diamine)(MTF-319)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-ethoxybenzimidamide (339 mg,1 mmol) to afford the titled compound as a white powder (291 mg, 86%).¹H NMR (200 MHz, DMSO-d6): δ 9.24 (s, 2H), 7.83 (d, J=8.8 Hz, 4H), 7.62(dd, J=7.6, 1.1 Hz, 2H), 7.37 (t, J=7.8 Hz, 4H), 7.32-7.23 (m, 2H),7.22-7.15 (m, 2H), 7.14-6.98 (m, 6H), 4.04 (q, J=6.9 Hz, 4H), 1.33 (t,J=6.9 Hz, 6H). ¹³C NMR (50 MHz, DMSO-d6): δ 169.98, 167.28, 165.48,158.46, 138.12, 136.73, 133.68, 129.30, 128.39, 127.64, 126.61, 126.29,120.13, 117.81, 113.27, 63.06, 14.68. HRMS-ESI (m/z): [M+H]⁺ calc. forC₃₄H₃₃N₁₀O₂S₂ ⁺, 677.22239; Found: 677.22253. HPLC (λ₂₈₀): Purity100.0%; t_(R): 13.058 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(3-fluorophenyl)-1,3,5-triazine-2,4-diamine(MTF-320)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-fluorobenzimidamide (313 mg,1 mmol) to afford the titled compound as a white powder (237 mg, 76%).¹H NMR (200 MHz, DMSO-d6): δ 9.32 (s, 2H), 8.08 (d, J=7.6 Hz, 2H), 7.93(d, J=9.7 Hz, 2H), 7.62 (d, J=7.7 Hz, 2H), 7.57-7.45 (m, 2H), 7.45-7.06(m, 12H). ¹⁹F NMR (188 MHz, DMSO-d6): δ −113.29. ¹³C NMR (50 MHz,DMSO-d6): δ 168.98, 167.26, 165.51, 164.58, 159.75, 139.31 (d, J=7.7Hz), 136.52, 133.76, 130.39 (d, J=7.8 Hz), 128.22, 127.66, 126.62 (d,J=16.1 Hz), 123.79, 118.23 (d, J=20.5 Hz), 114.11 (d, J=22.6 Hz).HRMS-ESI (m/z): [M+H]⁺ calc. for C₃₀H₂₃F₂N₁₀S₂ ⁺, 625.15112; Found:625.15125. HPLC (λ₂₈₀): Purity 97.5%; t_(R): 12.817 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(pyridin-2-yl)-1,3,5-triazine-2,4-diamine)(MTF-321)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)picolinimidamide (296 mg, 1mmol) to afford the titled compound as a white powder (266 mg, 90%). ¹HNMR (200 MHz, DMSO-d6): δ 9.39 (s, 1H), 8.74-8.64 (m, 1H), 8.20 (d,J=7.8 Hz, 1H), 7.93 (td, J=7.7, 1.8 Hz, 1H), 7.61 (dd, J=7.6, 1.5 Hz,1H), 7.51 (ddd, J=7.5, 4.7, 1.2 Hz, 1H), 7.39 (dd, J=7.7, 1.4 Hz, 1H),7.35-7.08 (m, 4H). ¹³C NMR (50 MHz, DMSO-d6): δ 170.30, 167.53, 165.79,154.32, 149.29, 136.74, 136.57, 133.61, 128.25, 127.64, 126.88, 126.47,125.48, 123.33. HRMS-ESI (m/z): [M+H]⁺ calc. for C₂₈H₂₃N₁₂S₂ ⁺,591.16048; Found: 591.16048. HPLC (λ₂₈₀): Purity 95.3%; t_(R): 6.483 min(method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(3-chlorophenyl)-1,3,5-triazine-2,4-diamine)(MTF-322)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-chlorobenzimidamide (329 mg,1 mmol) to afford the titled compound as a white powder (260 mg, 79%).¹H NMR (200 MHz, DMSO-d6): δ 9.33 (s, 2H), 8.24 (s, 2H), 8.16 (d, J=7.6Hz, 2H), 7.58 (ddd, J=20.7, 9.4, 4.5 Hz, 6H), 7.44-7.08 (m, 10H). ¹³CNMR (50 MHz, DMSO-d6): δ 168.82, 167.23, 165.50, 138.80, 136.49, 133.81,133.18, 131.14, 130.31, 128.22, 127.66, 127.45, 126.79, 126.51, 126.27.HRMS-ESI (m/z): [M+H]⁺ calc. for C₃₀H₂₃Cl₂N₁₀S₂ ⁺, 657.09201; Found:657.09222. HPLC (λ₂₈₀): Purity 96.9%; t_(R): 12.867 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(3-bromophenyl)-1,3,5-triazine-2,4-diamine)(MTF-323)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-bromobenzimidamide (374 mg,1 mmol) to afford the titled compound as a white powder (339 mg, 91%).¹H NMR (200 MHz, DMSO-d6): δ 9.34 (s, 2H), 8.40 (s, 2H), 8.20 (d, J=7.8Hz, 2H), 7.74 (ddd, J=8.0, 2.0, 1.0 Hz, 2H), 7.62 (dd, J=7.5, 1.6 Hz,2H), 7.46 (t, J=7.9 Hz, 2H), 7.40-7.13 (m, 10H). ¹³C NMR (50 MHz,DMSO-d6): δ 168.73, 167.22, 165.49, 138.97, 136.48, 134.02, 133.83,130.60, 130.43, 128.22, 127.66, 126.81, 126.63, 126.55, 121.69. HRMS-ESI(m/z): [M+H]⁺ calc. for C₃₀H₂₃Br₂N₁₀S₂ ⁺, 744.99098; Found: 744.99098.HPLC (λ₂₈₀): Purity 100.0%; t_(R): 17.600 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(4-methoxyphenyl)-1,3,5-triazine-2,4-diamine)(MTF-324)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-methoxybenzimidamide (325mg, 1 mmol) to afford the titled compound as a white powder (263 mg,81%). ¹H NMR (200 MHz, DMSO-d6): δ 9.13 (s, 2H), 8.20 (d, J=8.8 Hz, 4H),7.62 (dd, J=7.8, 1.2 Hz, 2H), 7.40 (d, J=7.2 Hz, 2H), 7.32-7.22 (m, 2H),7.22-7.12 (m, 2H), 7.01 (d, J=8.8 Hz, 8H), 3.82 (s, 6H). ¹³C NMR (50MHz, DMSO-d6): δ 169.77, 167.19, 165.36, 161.99, 136.91, 133.33, 129.62(2C), 128.90, 128.61, 127.72, 126.36, 126.10, 113.57 (2C), 55.30.HRMS-ESI (m/z): [M+H]⁺ calc. for C₃₂H₂₉N₁₀O₂S₂ ⁺, 649.19109; Found:649.19116. HPLC (λ₂₈₀): Purity 100.0%; t_(R): 10.800 min (method 2).

3,3′-(((disulfanediylbis(2,1-phenylene))bis(azanediyl))bis(6-amino-1,3,5-triazine-4,2-diyl))dibenzonitrile(MTF-325)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-cyanobenzimidamide (320 mg,1 mmol) to afford the titled compound as a white powder (291 mg, 91%).¹H NMR (200 MHz, DMSO-d6): δ 9.37 (s, 2H), 8.49 (d, J=10.9 Hz, 4H), 8.02(d, J=7.7 Hz, 2H), 7.72 (t, J=7.8 Hz, 2H), 7.63 (dd, J=7.6, 1.4 Hz, 2H),7.42-7.15 (m, 10H). ¹³C NMR (50 MHz, DMSO-d6): δ 168.32, 167.24, 165.49,137.79, 136.48, 134.74, 133.71, 132.16, 131.22, 129.88, 128.38, 127.79,126.80, 126.59, 118.56, 111.54. HRMS-ESI (m/z): [M+H]⁺ calc. forC₃₂H₂₃N₁₂S₂ ⁺, 639.16046; Found: 639.16064. HPLC (λ₂₈₀): Purity 96.1%;t_(R): 11.858 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-phenyl-1,3,5-triazine-2,4-diamine)(MTF-326)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)benzimidamide (295 mg, 1 mmol)to afford the titled compound as a white powder (218 mg, 74%). ¹H NMR(200 MHz, DMSO-d6): δ 9.23 (s, 2H), 8.24 (d, J=6.5 Hz, 4H), 7.62 (d,J=7.3 Hz, 2H), 7.57-7.34 (m, 8H), 7.33-7.01 (m, 8H). ¹³C NMR (50 MHz,DMSO-d6): δ 170.16, 167.29, 165.51, 136.71, 136.61, 133.59, 131.41,128.37, 128.24 (2C), 127.82 (2C), 127.66, 126.61, 126.31. HRMS-ESI(m/z): [M+H]⁺ calc. for C₃₀H₂₅N₁₀S₂ ⁺, 589.16996; Found: 589.16992. HPLC(λ₂₈₀): Purity 100.0%; t_(R): 11.792 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(4-chlorophenyl)-1,3,5-triazine-2,4-diamine)(MTF-327)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-chlorobenzimidamide (329 mg,1 mmol) to afford the titled compound as a white powder (283 mg, 86%).¹H NMR (200 MHz, DMSO-d6): δ 9.27 (s, 2H), 8.22 (d, J=8.5 Hz, 4H),7.66-7.50 (m, 6H), 7.38 (dd, J=7.6, 1.0 Hz, 2H), 7.33-7.05 (m, 8H). ¹³CNMR (50 MHz, DMSO-d6): δ 169.18, 167.22, 165.46, 136.62, 136.20, 135.47,133.52, 129.53 (2C), 128.42 (3C), 127.73, 126.65, 126.37. HRMS-ESI(m/z): [M+H]⁺ calc. for C₃₀H₂₃O₂N₁₀S₂ ⁺, 657.09201; Found: 657.09210.HPLC (λ₂₈₀): Purity 100.0%; t_(R): 15.633 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(4-iodophenyl)-1,3,5-triazine-2,4-diamine)(MTF-328)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-iodobenzimidamide (421 mg, 1mmol) to afford the titled compound as a white powder (387 mg, 92%). ¹HNMR (200 MHz, DMSO-d6): δ 9.26 (s, 2H), 8.00 (d, J=8.4 Hz, 4H), 7.87 (d,J=8.4 Hz, 4H), 7.61 (d, J=7.6 Hz, 2H), 7.39 (d, J=7.4 Hz, 2H), 7.33-6.99(m, 8H). ¹³C NMR (50 MHz, DMSO-d6): δ 169.58, 167.21, 165.45, 137.24(2C), 136.63, 136.19, 133.49, 129.70 (2C), 128.47, 127.73, 126.61,126.34, 99.15. HRMS-ESI (m/z): [M+H]⁺ calc. for C₃₀H₂₃I₂N₁₀S₂ ⁺,840.96324; Found: 840.96289. HPLC (λ₂₈₀): Purity 100.0%; t_(R): 17.833min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(pyridin-4-yl)-1,3,5-triazine-2,4-diamine)(MTF-329)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)isonicotinimidamide (296 mg, 1mmol) to afford the titled compound as a white powder (266 mg, 90%). ¹HNMR (200 MHz, DMSO-d6): δ 9.41 (s, 2H), 8.73 (dd, J=4.5, 1.5 Hz, 4H),8.05 (d, J=5.8 Hz, 4H), 7.62 (dd, J=7.4, 1.2 Hz, 2H), 7.43-7.08 (m,10H). ¹³C NMR (50 MHz, DMSO-d6): δ 168.77, 167.33, 165.59, 150.23 (2C),144.04, 136.41, 133.74, 128.33, 127.75, 126.86, 126.61, 121.55 (2C).HRMS-ESI (m/z): [M+H]⁺ calc. for C₂₈H₂₃N₁₂S₂ ⁺, 591.16046; Found:591.16089. HPLC (λ₂₈₀): Purity 99.2%; t_(R): 7.742 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(pyridin-3-yl)-1,3,5-triazine-2,4-diamine)(MTF-330)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)nicotinimidamide (296 mg, 1mmol) to afford the titled compound as a white powder (266 mg, 90%).(277 mg, 94%). ¹H NMR (200 MHz, DMSO-d6): δ 9.35 (s, 4H), 8.71 (dd,J=4.8, 1.7 Hz, 2H), 8.47 (d, J=8.1 Hz, 2H), 7.62 (dd, J=7.6, 1.2 Hz,2H), 7.52 (dd, J=7.8, 4.6 Hz, 2H), 7.38 (dd, J=7.7, 1.4 Hz, 2H),7.33-7.06 (m, 8H¹³C NMR (50 MHz, DMSO-d6): δ 168.81, 167.12, 165.37,151.99, 149.09, 136.50, 135.09, 133.66, 132.00, 128.31, 127.69, 126.77,126.49, 123.51 HRMS-ESI (m/z): [M+H]⁺ calc. for C₂₈H₂₃N₁₂S₂ ⁺,591.16046; Found: 591.16052. HPLC (λ₂₈₀): Purity 100.0%; t_(R): 7.700min (method 2).

2-((4-amino-6-(3-bromophenyl)-1,3,5-triazin-2-yl)amino)phenol (MTF-331)

Synthesized following the general procedure A using1-(benzo[d]oxazol-2-yl)guanidine (1.00 g, 5.68 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 341 mg, 8.52 mmol) and3-bromobenzonitrile (1.034 g, 5.68 mmol) to afford the titled compoundas a brown powder (1.46 g, 72%). ¹H NMR (400 MHz, DMSO-d6): δ 9.97 (s,1H), 8.44 (s, 1H), 8.36 (s, 1H), 8.26 (d, J=7.5 Hz, 1H), 7.90 (d, J=7.7Hz, 1H), 7.76 (d, J=7.7 Hz, 1H), 7.48 (t, J=7.8 Hz, 1H), 7.29 (s, 2H),6.99-6.92 (m, 1H), 6.89 (d, J=7.4 Hz, 1H), 6.82 (t, J=7.1 Hz, 1H). ¹³CNMR (50 MHz, DMSO-d6): δ 168.84, 167.06, 164.48, 148.30, 138.75, 133.26,131.22, 130.38, 127.46, 126.84, 126.29, 124.12, 122.65, 118.99, 115.77.HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₅H₁₃BrN₅O⁺, 358.02980 Found:358.03094. HPLC (λ₂₈₀): Purity 97.1%; t_(R): 9.808 min (method 1).

2-((4-amino-6-(3-chlorophenyl)-1,3,5-triazin-2-yl)amino)phenol (MTF-332)

Synthesized following the general procedure A using1-(benzo[d]oxazol-2-yl)guanidine (1.00 g, 5.68 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 341 mg, 8.52 mmol) and3-chlorobenzonitrile (781 mg, 5.68 mmol) to afford the titled compoundas a brown powder (1.45 g, 82%). ¹H NMR (400 MHz, DMSO-d6): δ 10.03 (s,1H), 8.36 (s, 1H), 8.29 (s, 1H), 8.23 (d, J=7.4 Hz, 1H), 7.92 (d, J=7.5Hz, 1H), 7.62 (d, J=7.5 Hz, 1H), 7.55 (t, J=7.8 Hz, 1H), 7.29 (s, 2H),6.99-6.92 (m, 1H), 6.89 (d, J=7.3 Hz, 1H), 6.82 (t, J=7.1 Hz, 1H). ¹³CNMR (50 MHz, DMSO-d6): δ 168.74, 167.02, 164.48, 148.21, 138.90, 134.11,130.67, 130.40, 126.66 (2C), 124.17, 122.79, 121.73, 119.10, 115.78.HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₅H₁₃ClN₅O⁺, 314.08031; Found:314.08066. HPLC (λ₂₈₀): Purity 97.3%; t_(R): 9.625 min (method 1).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-benzyl-1,3,5-triazine-2,4-diamine(MTF-333)

In a microwave tube, 1-(benzo[d]thiazol-2-yl)guanidine (100 mg, 0.52mmol) was solubilized in NMP (2 mL) under argon atmosphere. The tube wassubsequently cooled down to 0° C. before NaH (60% suspension in oil, 23mg, 0.57 mmol) was added. After the gas evolution stopped, the tube wassealed and the mixture was warmed up to 110° C. under microwaveirradiation for 15 min. The resulting slurry was taken in Et₂O andfiltered. The precipitate was then purified by silicagel flashchromatography (dichloromethane/MeOH, 10/0 to 9/1) to give the desiredproduct as a white powder (50 mg, 31%). TLC: R_(f)(dichloromethane/MeOH, 9/1, v/v)=0.83. ¹H NMR (400 MHz, DMSO-d6): δ 9.05(s, 1H), 7.54 (dd, J=7.8, 0.9 Hz, 1H), 7.35-7.17 (m, 7H), 7.14 (t, J=7.7Hz, 1H), 6.95 (d, J=16.0 Hz, 2H), 3.71 (s, 2H). ¹³C NMR (101 MHz,DMSO-d6): δ 176.6, 166.9, 165.3, 151.5, 139.2, 137.9, 136.6, 129.1,128.2, 128.0, 127.6, 126.3, 124.9, 30.4. HRMS-ESI (m/z): [M+H]⁺ calc.for C₃₂H₂₉N₁₀S₂ ⁺617.20126, found 617.20154. HPLC (λ₂₅₄): Purity 96.2%;t_(R): 10.742 min (method 2).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-methoxybenzimidamide(MTF-342)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and3-methoxybenzonitrile (0.635 mL, 5.20 mmol) to afford the titledcompound as a white powder (914 mg, 54%). ¹H NMR (200 MHz, DMSO-d6): δ10.26 (s, 1H), 9.33 (s, 1H), 8.82 (s, 1H), 8.05 (s, 1H), 7.80 (d, J=7.6Hz, 1H), 7.73-7.52 (m, 3H), 7.48-7.29 (m, 2H), 7.17 (dd, J=14.6, 7.6 Hz,2H), 3.83 (s, 3H). ¹³C NMR spectrum could not be properly recorded asthis compound rearranges and dimerizes into compound MTF-318 during thetime of the analysis. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₆H₁₆N₅OS⁺,326.10701; Found: 326.10706. HPLC (λ₂₈₀): Purity 100.0%; t_(R): 6.558min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-(trifluoromethyl)benzimidamide(MTF-343)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and3-(trifluoromethyl)benzonitrile (890 mg, 5.20 mmol) to afford the titledcompound as a white powder (435 mg, 23%). ¹H NMR (200 MHz, DMSO-d6): δ10.33 (br. s, 1H), 9.36 (br. s, 1H), 8.93 (br. s, 1H), 8.39 (s, 1H),8.31 (d, J=7.9 Hz, 1H), 8.16 (br. s, 1H), 7.95 (d, J=7.9 Hz, 1H),7.86-7.72 (m, 2H), 7.67 (d, J=8.0 Hz, 1H), 7.35 (t, J=7.6 Hz, 1H), 7.20(t, J=7.5 Hz, 1H). ¹⁹F NMR (376 MHz, DMSO-d6): δ −61.04. ¹³C NMRspectrum could not be properly recorded as this compound rearranges anddimerizes during the time of the analysis. HRMS-ESI (m/z): [M+H]⁺ calc.for C₁₆H₁₃F₃N₅S⁺, 364.08383; Found: 364.08408. HPLC (λ₂₈₀): Purity97.9%; t_(R): 6.533 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-ethoxybenzimidamide(MTF-344)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and3-ethoxybenzonitrile (765 mg, 5.20 mmol) to afford the titled compoundas a white powder (830 mg, 47%). ¹H NMR (200 MHz, DMSO-d6): δ 10.23 (br.s, 1H), 9.30 (br. s, 1H), 8.78 (br. s, 1H), 8.03 (br. s, 1H), 7.80 (dd,J=7.8, 0.8 Hz, 1H), 7.65 (dd, J=8.0, 0.6 Hz, 1H), 7.62-7.53 (m, 2H),7.46-7.29 (m, 2H), 7.24-7.07 (m, 2H), 4.09 (q, J=6.9 Hz, 2H), 1.36 (t,J=6.9 Hz, 3H). ¹³C NMR (50 MHz, DMSO-d6): δ 172.24, 162.07, 161.82,158.40, 151.49, 136.56, 131.16, 129.36, 125.63, 122.78, 121.13, 119.68,119.65, 117.34, 113.64, 63.24, 14.63. HRMS-ESI (m/z): [M+H]⁺ calc. forC₁₇H₁₈N₅OS⁺, 340.12266; Found: 340.12296. HPLC (λ₂₈₀): Purity 100.0%;t_(R): 6.733 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)pyrazine-2-carboximidamide(MTF-345)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) andpyrazinecarbonitrile (0.465 mL, 5.20 mmol) to afford the titled compoundas a yellow powder (959 mg, 62%). ¹H NMR (200 MHz, DMSO-d6): δ 10.13(br. s, 1H), 9.50 (s, 1H), 9.44 (br. s, 1H), 8.99 (br. s, 1H), 8.88 (d,J=2.5 Hz, 1H), 8.79 (dd, J=2.5, 1.4 Hz, 1H), 8.26 (br. s, 1H), 7.83 (d,J=7.7 Hz, 1H), 7.69 (d, J=7.9 Hz, 1H), 7.44-7.30 (m, 1H), 7.29-7.15 (m,1H). ¹³C NMR (50 MHz, DMSO-d6): δ 172.18, 161.92, 157.68, 151.39,147.21, 145.85, 143.84, 143.44, 131.22, 125.73, 123.01, 121.22, 119.90.HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₃H₁₂N₇S⁺, 298.08694; Found:298.08707. HPLC (λ₂₈₀): Purity 95.7%; t_(R): 5.508 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-fluorobenzimidamide(MTF-346)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and3-fluorobenzonitrile (0.56 mL, 5.20 mmol) to afford the titled compoundas a white powder (895 mg, 55%). ¹H NMR (200 MHz, DMSO-d6): δ 10.26 (s,1H), 9.34 (s, 1H), 8.89 (s, 1H), 8.09 (s, 1H), 7.95-7.76 (m, 3H),7.70-7.51 (m, 2H), 7.50-7.28 (m, 2H), 7.19 (td, J=7.6, 1.2 Hz, 1H). ¹⁹FNMR (376 MHz, DMSO-d6): δ −112.99. ¹³C NMR spectrum could not beproperly recorded as this compound rearranges and dimerizes intocompound MTF-320 during the time of the analysis. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₅H₁₃FN₅S⁺, 314.08702; Found: 314.08722. HPLC (λ₂₈₀): Purity98.7%; t_(R): 6.442 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-cyanobenzimidamide (MTF-347)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and1,3-dicyanobenzene (667 mg, 5.20 mmol) to afford the titled compound asa white-yellowish powder (1.23 g, 74%). ¹H NMR (200 MHz, DMSO-d6): δ10.27 (br. s, 1H), 9.37 (br. s, 1H), 8.97 (br. s, 1H), 8.45 (s, 1H),8.33 (dd, J=8.0, 1.0 Hz, 1H), 8.15 (br. s, 1H), 8.06 (dd, J=7.7, 1.0 Hz,1H), 7.89-7.62 (m, 3H), 7.35 (t, J=7.6 Hz, 1H), 7.20 (t, J=7.5 Hz, 1H).¹³C NMR (50 MHz, DMSO-d6): δ 172.17, 161.83, 159.93, 151.42, 136.30,134.69, 132.15, 131.40, 131.24, 129.67, 125.67, 122.89, 121.17, 119.81,118.39, 111.50. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₆H₁₃N₆S⁺, 321.09169;Found: 321.09167. HPLC (λ₂₈₀): Purity 96.4%; t_(R): 6.342 min (method1).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(pyrazin-2-yl)-1,3,5-triazine-2,4-diamine)(MTF-348)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)pyrazine-2-carboximidamide (297mg, 1 mmol) to afford the titled compound as a yellowish powder (279 mg,94%). ¹H NMR (400 MHz, DMSO-d6): δ 9.48 (br. s, 1H), 9.33 (br. s, 1H),8.77 (m, 2H), 7.61 (d, J=7.7 Hz, 1H), 7.38 (d, J=6.7 Hz, 2H), 7.23 (m,3H). ¹³C NMR (50 MHz, DMSO-d6): δ 168.83, 167.37, 165.65, 149.51,146.27, 144.58, 144.38, 136.35, 133.76, 128.18, 127.66, 127.06, 126.68.HRMS-ESI (m/z): [M+H]⁺ calc. for C₂₈H₂₃N₁₂S₂ ⁺, 591.16046; Found:591.16052. HPLC (λ₂₈₀): Purity 99.2%; t_(R): 8.875 min (method 2).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-6-chloronicotinimidamide(MTF-379)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and6-chloro-3-pyridinecarbonitrile (720 mg, 5.20 mmol) to afford the titledcompound as a beige powder (1.48 g, 86%). ¹H NMR (400 MHz, DMSO-d6): δ10.21 (s, 1H), 9.37 (s, 1H), 9.01 (d and br. s, J=2.1 Hz, 2H), 8.38 (dd,J=8.4, 2.3 Hz, 1H), 8.12 (s, 1H), 7.80 (d, J=7.8 Hz, 1H), 7.69 (d, J=8.4Hz, 1H), 7.66 (d, J=8.1 Hz, 1H), 7.34 (t, J=7.6 Hz, 1H), 7.19 (t, J=7.5Hz, 1H). ¹³C NMR (101 MHz, DMSO-d6): δ 172.16, 161.79, 159.12, 152.70,151.42, 149.28, 138.70, 131.29, 130.36, 125.68, 123.98, 122.91, 121.17,119.83. HPLC (λ₂₈₀): Purity 97.2%; t_(R): 7.033 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2-chloroisonicotinimidamide(MTF-380)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and2-chloro-4-pyridinecarbonitrile (720 mg, 5.20 mmol) to afford the titledcompound as a beige powder (327 mg, 19%). ¹H NMR (400 MHz, DMSO-d6): δ10.22 (br. s, 1H), 9.39 (br. s, 1H), 9.04 (br. s, 1H), 8.60 (d, J=5.1Hz, 1H), 8.21 (br. s, 1H), 8.05 (s, 1H), 7.94 (dd, J=5.2, 1.3 Hz, 1H),7.81 (d, J=7.3 Hz, 1H), 7.67 (d, J=7.9 Hz, 1H), 7.38-7.31 (m, 1H),7.24-7.17 (m, 1H). ¹³C NMR (101 MHz, DMSO-d6): δ 172.08, 161.68, 158.50,151.36, 150.84, 150.52, 146.10, 131.31, 125.70, 122.99, 122.27, 121.20,120.99, 119.90. HPLC (λ₂₈₀): Purity 97.8%; t_(R): 7.108 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)thiophene-2-carboximidamide(MTF-381)

Synthesized following the general procedure1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and2-thiophenecarbonitrile (0.48 mL, 5.20 mmol) to afford the titledcompound as a yellowish powder (815 mg, 52%). ¹H NMR (400 MHz, DMSO-d6):δ 10.22 (br. s, 1H), 9.34 (br. s, 1H), 8.88 (br. s, 1H), 7.94 (dd,J=3.7, 0.9 Hz, 2H), 7.79 (d, J=6.2 Hz, 2H), 7.65 (d, J=7.9 Hz, 1H),7.39-7.28 (m, 1H), 7.24-7.14 (m, 2H). ¹³C NMR (101 MHz, DMSO-d6): δ172.22, 161.71, 157.31, 151.46, 140.40, 131.89, 131.13, 128.97, 128.11,125.65, 122.79, 121.12, 119.68. HPLC (λ₂₈₀): Purity 99.2%; t_(R): 6.975min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3,4,5-trimethoxybenzimidamide(MTF-382)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and3,4,5-trimethoxybenzonitrile (1 g, 5.20 mmol) to afford the titledcompound as a beige powder (441 mg, 22%). ¹H NMR (200 MHz, DMSO-d6): δ10.22 (br. s, 1H), 9.29 (br. s, 1H), 8.77 (br. s, 1H), 8.01 (br. s, 1H),7.80 (d, J=7.0 Hz, 1H), 7.65 (d, J=7.5 Hz, 1H), 7.39 (s, 2H), 7.38-7.27(m, 1H), 7.19 (td, J=7.7, 1.1 Hz, 1H), 3.87 (s, 6H), 3.74 (s, 3H). ¹³CNMR (50 MHz, DMSO-d6): δ 172.28, 161.97, 161.53, 152.51 (2C), 151.54,140.27, 131.19, 130.33, 125.63, 122.75, 121.12, 119.66, 105.29 (2C),60.14, 56.06 (2C). HPLC (λ₂₈₀): Purity 96.1%; t_(R): 7.175 min (method1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)benzo[d][1,3]dioxole-5-carboximidamide(MTF-383)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) andpiperonylonitrile (765 mg, 5.20 mmol) to afford the titled compound as abeige powder (600 mg, 34%). ¹H NMR (400 MHz, DMSO-d6): δ 10.27 (s, 1H),9.29 (s, 1H), 8.68 (s, 1H), 7.95 (s, 1H), 7.79 (d, J=7.7 Hz, 1H),7.68-7.62 (m, 2H), 7.60 (d, J=1.6 Hz, 1H), 7.37-7.29 (m, 1H), 7.22-7.15(m, 1H), 7.04 (d, J=8.2 Hz, 1H), 6.13 (s, 2H). ¹³C NMR (50 MHz,DMSO-d6): δ 172.25, 161.95, 161.27, 151.52, 150.06, 147.38, 131.13,128.99, 125.62, 122.74, 122.56, 121.11, 119.65, 107.85, 107.61, 101.80.HPLC (λ₂₈₀): Purity 96.0%; t_(R): 7.133 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2-naphthimidamide (MTF-384)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) andnaphthalene-2-carbonitrile (796 mg, 5.20 mmol) to afford the titledcompound as a beige powder (1.04 g, 58%). ¹H NMR (400 MHz, DMSO-d6): δ10.38 (br. s, 1H), 9.42 (br. s, 1H), 8.99 (br. s, 1H), 8.64 (s, 1H),8.16 (br. s and dd, J=8.6, 1.4 Hz, 2H), 8.08-7.96 (m, 3H), 7.81 (d,J=7.7 Hz, 1H), 7.67 (d, J=7.9 Hz, 1H), 7.66-7.57 (m, 2H), 7.40-7.31 (m,1H), 7.25-7.15 (m, 1H). ¹³C NMR (50 MHz, DMSO-d6): δ 172.34, 162.16(2C), 151.53, 134.30, 132.66, 132.15, 131.22, 128.83, 127.84, 127.75(2C), 127.66, 126.80, 125.65, 124.55, 122.81, 121.14, 119.73. HPLC(λ₂₈₀): Purity 95.7%; t_(R): 7.392 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-(trifluoromethyl)benzimidamide(MTF-385)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and4-(trifluoromethyl)benzonitrile (889 mg, 5.20 mmol) to afford the titledcompound as a beige powder (1.38 g, 73%). ¹H NMR (400 MHz, DMSO-d6): δ10.28 (br. s, 1H), 9.38 (br. s, 1H), 8.97 (br. s, 1H), 8.22 (d, J=8.2Hz, 2H), 8.11 (br. s, 1H), 7.90 (d, J=8.3 Hz, 2H), 7.80 (d, J=7.3 Hz,1H), 7.67 (d, J=7.7 Hz, 1H), 7.39-7.30 (m, 1H), 7.24-7.16 (m, 1H). ¹⁹FNMR (377 MHz, DMSO-d6): δ −61.28. ¹³C NMR (50 MHz, DMSO-d6): δ 172.11,161.89, 160.69, 151.37, 139.09, 131.19, 131.40 (q, J=31.8 Hz), 128.36(2C), 125.57, 125.16 (q, J=3.8 Hz, 2C), 123.91 (q, J=67.7 Hz), 122.79,121.07, 119.72. HPLC (λ₂₈₀): Purity 96.4%; t_(R): 7.442 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2-bromoisonicotinimidamide(MTF-386)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and2-bromo-4-cyanopyridine (952 mg, 5.20 mmol) to afford the titledcompound as a yellow powder (859 mg, 44%). ¹H NMR (400 MHz, DMSO-d6): δ10.19 (br. s, 1H), 9.38 (br. s, 1H), 9.04 (br. s, 1H), 8.58 (d, J=5.1Hz, 1H), 8.18 (s and br. s, 2H), 7.96 (dd, J=5.1, 1.3 Hz, 1H), 7.81 (d,J=7.4 Hz, 1H), 7.67 (d, J=7.9 Hz, 1H), 7.39-7.32 (m, 1H), 7.24-7.17 (m,1H). ¹³C NMR (50 MHz, DMSO-d6): δ 172.08, 161.67, 158.42, 151.36,151.00, 145.67, 141.81, 131.32, 125.94, 125.71, 123.00, 121.25, 121.21,119.91. HPLC (λ₂₈₀): Purity 97.0%; t_(R): 7.192 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-bromothiophene-3-carboximidamide(MTF-387)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and4-bromothiophene-3-carbonitrile (978 mg, 5.20 mmol) to afford the titledcompound as a brown powder (969 mg, 49%). ¹H NMR (400 MHz, DMSO-d6): δ10.03 (br. s, 1H), 9.30 (br. s, 1H), 8.60 (br. s, 1H), 7.98 (br.s and d,J=3.4 Hz, 2H), 7.80 (d, J=7.7 Hz, 1H), 7.77 (d, J=3.4 Hz, 1H), 7.64 (d,J=8.0 Hz, 1H), 7.38-7.30 (m, 1H), 7.20 (d, J=7.3 Hz, 1H). ¹³C NMR (50MHz, DMSO-d6): δ 172.41, 161.84, 159.08, 151.48, 137.36, 131.24, 129.06,125.66, 125.44, 122.85, 121.16, 119.75, 108.70. HPLC (λ₂₈₀): Purity96.6%; t_(R): 6.967 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2,2-dimethyl-2H-chromene-6-carboximidamide(MTF-388)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and2,2-dimethyl-2H-1-benzopyran-6-carbonitrile (963 mg, 5.20 mmol) toafford the titled compound as a beige powder (118 mg, 6%). ¹H NMR (400MHz, DMSO-d6): δ 10.25 (br. s, 1H), 9.29 (br. s, 1H), 8.66 (br. s, 1H),7.91 (br. s, 1H), 7.84-7.75 (m, 3H), 7.64 (d, J=7.8 Hz, 1H), 7.37-7.30(m, 1H), 7.22-7.15 (m, 1H), 6.85 (d, J=8.5 Hz, 1H), 6.45 (d, J=9.9 Hz,1H), 5.84 (d, J=9.8 Hz, 1H), 1.41 (s, 6H). ¹³C NMR (101 MHz, DMSO-d6): δ169.68, 167.16, 165.32, 155.42, 136.88, 133.37, 131.25, 129.27, 129.18,128.53, 127.69, 126.40, 126.16, 126.10, 121.61, 120.27, 115.66, 76.97,27.93 (2C). HPLC (λ₂₈₀): Purity 98.4%; t_(R): 7.492 min (method 1).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3,5-dichloropicolinimidamide(MTF-389)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and3,5-dichloropyridine-2-carbonitrile (900 mg, 5.20 mmol) to afford thetitled compound as a brown powder (779 mg, 41%). ¹H NMR (400 MHz,DMSO-d6): δ 9.73 (br. s, 1H), 9.29 (br. s, 1H), 8.73 (br. s, 1H), 8.67(d, J=1.5 Hz, 1H), 8.37 (s, 1H), 8.12 (br. s, 1H), 7.77 (d, J=7.7 Hz,1H), 7.62 (d, J=7.9 Hz, 1H), 7.32 (t, J=7.5 Hz, 1H), 7.18 (t, J=7.5 Hz,1H). ¹³C NMR (50 MHz, DMSO-d6): δ 172.48, 161.60, 160.60, 151.43,150.95, 145.79, 137.15, 131.46, 131.19, 129.02, 125.59, 122.77, 121.12,119.67. HPLC (λ₂₈₀): Purity 95.3%; t_(R): 7.192 min (method 1).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(4-(trifluoromethyl)phenyl)-1,3,5-triazine-2,4-diamine)(MTF-394)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-(trifluoromethyl)benzimidamide(363 mg, 1 mmol) to afford the titled compound as a white powder (239mg, 66%). ¹H NMR (400 MHz, DMSO-d6): δ 9.34 (s, 1H), 8.42 (d, J=7.8 Hz,2H), 7.86 (d, J=8.4 Hz, 2H), 7.64 (d, J=7.7 Hz, 1H), 7.41 (d, J=7.4 Hz,1H), 7.25 (ddd, J=34.2, 11.4, 4.1 Hz, 4H). ¹⁹F NMR (377 MHz, DMSO-d6): δ−61.24. ¹³C NMR (101 MHz, DMSO-d6): δ 168.98, 167.28, 165.53, 140.54,136.56, 133.57, 131.18, 128.46 (2C), 127.76, 126.75, 126.48, 125.50,125.30 (2C), 122.80. HPLC (λ₂₅₄): Purity 97.0%; t_(R): 16.125 min(method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(thiophen-2-yl)-1,3,5-triazine-2,4-diamine)(MTF-396)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)thiophene-2-carboximidamide(301 mg, 1 mmol) to afford the titled compound as a white powder (250mg, 83%). ¹H NMR (400 MHz, DMSO-d6): δ 9.21 (s, 1H), 7.84 (d, J=2.9 Hz,1H), 7.75 (dd, J=5.0, 1.1 Hz, 1H), 7.61 (dd, J=7.9, 1.1 Hz, 1H), 7.37(d, J=7.4 Hz, 1H), 7.26 (td, J=7.7, 1.2 Hz, 1H), 7.21-7.16 (m, 2H), 7.09(d, J=15.2 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d6): δ 166.91, 166.58,165.13, 142.40, 136.55, 133.67, 131.03, 129.34, 128.37, 128.08, 127.60,126.68, 126.35. HPLC (λ₂₈₀): Purity 97.1%; t_(R): 11.817 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(4-bromothiophen-3-yl)-1,3,5-triazine-2,4-diamine)(MTF-397)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-bromothiophene-3-carboximidamide(380 mg, 1 mmol) to afford the titled compound as a white powder (167mg, 44%). ¹H NMR (400 MHz, DMSO-d6): δ 9.14 (s, 1H), 8.11 (d, J=3.6 Hz,1H), 7.73 (d, J=3.6 Hz, 1H), 7.57 (d, J=7.3 Hz, 1H), 7.42 (d, J=7.7 Hz,1H), 7.28-7.22 (m, 1H), 7.12 (t, J=7.4 Hz, 1H), 7.07 (s, 2H). ¹³C NMR(101 MHz, DMSO-d6): δ 167.68, 166.88, 165.20, 137.92, 136.60, 133.26,130.88, 128.44, 127.69, 126.98, 126.29, 126.03, 109.03. HPLC (λ₂₅₄):Purity 97.1%; t_(R): 12.483 min (method 2).

N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-nitrobenzimidamide (MTF-398)

Synthesized following the general procedure A using1-(benzo[d]thiazol-2-yl)guanidine (1.00 g, 5.20 mmol), sodium hydride(60% dispersion in mineral oil, 1.5 eq., 312 mg, 7.81 mmol) and4-nitrobenzonitrile (770 mg, 5.20 mmol) to afford the titled compound asa yellow powder (212 mg, 12%). ¹H NMR (500 MHz, DMSO-d6): δ 10.30 (br.s, 1H), 9.37 (br. s, 1H), 8.90 (br. s, 1H), 8.12 (t, br. s, J=1.8 Hz,2H), 7.99 (d, J=7.9 Hz, 1H), 7.80 (d, J=7.7 Hz, 1H), 7.66 (d, J=7.9 Hz,1H), 7.65-7.60 (m, 1H), 7.54 (t, J=7.9 Hz, 1H), 7.38-7.30 (m, 1H),7.22-7.17 (m, 1H). ¹³C NMR (50 MHz, DMSO-d6): δ 172.22, 161.95, 160.57,151.47, 137.26, 133.22, 131.23, 131.20, 130.20, 127.53, 126.14, 125.67,122.87, 121.16, 119.79. HPLC (λ₂₈₀): Purity 99.2%; t_(R): 6.792 min(method 1).

2-((4-amino-6-(trichloromethyl)-1,3,5-triazin-2-yl)amino)phenol (MTF373)

Synthesized following the general procedure D using1-(benzo[d]oxazol-2-yl)guanidine (100 mg, 0.6 mmol) andtrichloroacetonitrile (600 μL, 6 mmol) to afford the title compound as abeige powder (61 mg, 32%). ¹H NMR (400 MHz, Acetone-d6): δ 9.10 (s, 1H),8.38 (br. s, 1H), 7.99 (br. s, 1H), 7.28 (br. s, 1H), 7.15 (br. s, 1H),7.05-6.92 (m, 2H), 6.87 (td, J=7.8, 1.7 Hz, 1H). ¹³C NMR (101 MHz,Acetone-d6): δ 174.0, 168.9, 165.8, 148.6, 127.6, 125.6, 123.2, 120.8,117.2, 97.4. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₀H₉ON₅Cl₃ ⁺319.98672;Found 319.98709. HPLC (λ₂₅₄): 97.9%; t_(R): 9.108 min (method 4).

2-((4-amino-6-(trichloromethyl)-1,3,5-triazin-2-yl)amino)-4-chlorophenol(MTF374)

Synthesized following the general procedure D using2-(5-chlorobenzo[d]oxazol-2-yl)guanidine (100 mg, 0.48 mmol) andtrichloroacetonitrile (450 μL, 4.8 mmol) to afford the title compound asa light pink powder (55 mg, 32%). ¹H NMR (400 MHz, Acetone-d6): δ 9.48(s, 1H), 8.30 (s, 1H), 8.18 (s, 1H), 7.43 (s, 1H), 7.20 (s, 1H),7.01-6.92 (m, 2H). ¹³C NMR (101 MHz, Acetone-d6): δ 174.0, 168.9, 165.6,146.6, 128.8, 124.9, 124.3, 121.9, 117.3, 97.3. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₀H₈ON₅Cl₄ ⁺353.94775; Found 353.94830. HPLC (λ₂₅₄): 97.1%;t_(R): 10.504 min (method 4).

2-(4-amino-6-(trichloromethyl)-1,3,5-triazin-2-yl)amino)-5-chlorophenol(MTF375)

Synthesized following the general procedure D using2-(6-chlorobenzo[d]oxazol-2-yl)guanidine (100 mg, 0.48 mmol) andtrichloroacetonitrile (480 μL, 4.8 mmol) to afford the title compound asa light green powder (59 mg, 35%). ¹H NMR (400 MHz, Acetone-d6): δ 9.66(br. s, 1H), 8.32 (br. s, 1H), 8.04 (br. s, 1H), 7.29 (br. s, 1H), 7.18(br. s, 1H), 6.99 (d, J=2.2 Hz, 1H), 6.90 (dd, J=8.7, 2.1 Hz, 1H). ¹³CNMR (101 MHz, Acetone-d6): δ 174.0, 168.9, 165.7, 149.5, 129.5, 126.7,124.1, 120.4, 116.8, 97.3. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₀H₈ON₅Cl₄⁺353.94775; Found 353.94837. HPLC (λ₂₅₄): 95.3%; t_(R): 10.615 min(method 4).

2-((4-amino-6-(trichloromethyl)-1,3,5-triazin-2-yl)amino)-5-nitrophenol(MTF376)

Synthesized following the general procedure D using2-(6-nitrobenzo[d]oxazol-2-yl)guanidine (100 mg, 0.45 mmol) andtrichloroacetonitrile (450 μL, 4.5 mmol) to afford the title compound asa light yellow powder (50 mg, 30%). ¹H NMR (400 MHz, MeOD): δ 8.72 (d,J=9.1 Hz, 1H), 7.78 (dd, J=9.1, 2.5 Hz, 1H), 7.70 (d, J=2.5 Hz, 1H). ¹³CNMR (101 MHz, Acetone-d6): δ 174.3, 169.1, 165.7, 147.1, 143.7, 134.5,120.3, 116.6, 110.4, 97.1. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₀H₈O₃N₆Cl₃⁺ 364.97180; Found 364.97208. HPLC (λ₂₅₄): 95.1%; t_(R): 10.623 min(method 4).

2-((4-amino-6-(trichloromethyl)-1,3,5-triazin-2-yl)amino)-4-nitrophenol(MTF377)

Synthesized following the general procedure D using2-(5-nitrobenzo[d]oxazol-2-yl)guanidine (100 mg, 0.45 mmol) andtrichloroacetonitrile (450 μL, 4.5 mmol) to afford the title compound asa light orange powder (117 mg, 71%). ¹H NMR (400 MHz, Acetone-d6): δ9.29 (br. s, 1H), 8.32 (br. s, 1H), 7.91 (dd, J=8.9, 2.6 Hz, 1H), 7.54(br. s, 1H), 7.23 (br. s, 1H), 7.11 (d, J=8.9 Hz, 1H), 6.61 (br. s, 1H).¹³C NMR (101 MHz, Acetone-d6): δ 174.3, 168.9, 165.6, 146.9, 143.6,134.3, 120.3, 116.7, 110.3, 97.1. HRMS-ESI (m/z): [M+H]⁺ calc. forC₁₀H₈O₃N₆Cl₃ ⁺364.97180; Found 364.97229. HPLC (λ₂₅₄): 97.7%; t_(R):9.434 min (method 4).

2-((4-amino-6-phenyl-1,3,5-triazin-2-yl)amino)-4-chlorophenol (MTF409)

Synthesized following the general procedure E using1-(5-chlorobenzo[d]oxazol-2-yl)guanidine (500 mg, 2.4 mmol), sodiumhydride (60% oil suspension, 104 mg, 2.6 mmol) and benzonitrile (0.25mL, 2.4 mmol) to afford the title compound as a beige powder (210 mg,28%). ¹H NMR (400 MHz, Acetone-d6): δ 9.88 (br. s, 1H), 8.41-8.37 (m,2H), 8.16 (s, 2H), 7.56 (t, J=7.2 Hz, 1H), 7.50 (t, J=7.3 Hz, 2H),7.01-6.92 (m, 2H), 6.81 (br. s, 2H). ¹³C NMR (101 MHz, Acetone-d6): δ172.1, 168.6, 165.7, 147.0, 137.3, 132.6, 129.9, 129.1 (2C), 129.0 (2C),124.8, 124.1, 121.9, 118.4. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₅H₁₃OClN₅⁺314.08031; Found 314.08044. HPLC (λ₂₅₄): 98.1%; t_(R): 9.916 min(method 4).

2-((6-imino-4-(trichloromethyl)-1,6-dihydro-1,3,5-triazin-2-yl)amino)-5-methylphenol(MTF410)

Synthesized following the general procedure D using1-(6-methylbenzo[d]oxazol-2-yl)guanidine (500 mg, 2.6 mmol) andtrichloroacetonitrile (2.6 mL, 26 mmol) to afford the title compound asa light grey powder (480 mg, 55%). ¹H NMR (400 MHz, Acetone-d6): δ 9.01(br. s, 1H), 8.31 (br. d, J=75.1 Hz, 1H), 7.79 (br. d, J=51.9 Hz, 1H),7.25 (br. s, 1H), 7.14 (br. s, 1H), 6.79 (d, J=0.9 Hz, 1H), 6.69 (dd,J=8.1, 1.0 Hz, 1H), 2.25 (s, 3H). ¹³C NMR (101 MHz, Acetone-d6): δ173.8, 168.7, 165.6, 149.0, 135.8, 124.7, 123.6, 121.3, 118.0, 97.3,20.9. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₁H₁₁OCl₃N₅ ⁺334.00237; Found334.00250. HPLC (λ₂₅₄): 100%; t_(R): 9.948 min (method 4).

2-((4-amino-6-phenyl-1,3,5-triazin-2-yl)amino)-5-methylphenol (MTF411)

Synthesized following the general procedure E using1-(6-methylbenzo[d]oxazol-2-yl)guanidine (500 mg, 2.6 mmol), sodiumhydride (60% oil suspension, 114 mg, 2.9 mmol) and benzonitrile (0.27mL, 2.6 mmol) to afford the title compound as a beige powder (410 mg,54%). ¹H NMR (400 MHz, Acetone-d6): δ 9.75 (br. s, 1H), 8.37 (d, J=7.4Hz, 2H), 8.30 (br. s, 1H), 7.59-7.53 (m, 2H), 7.48 (t, J=7.3 Hz, 2H),6.80 (s, 1H), 6.74 (br. s, 2H), 6.69 (d, J=8.0 Hz, 1H), 2.26 (s, 3H).¹³C NMR (101 MHz, Acetone-d6): δ 171.9, 168.4, 165.6, 149.1, 137.4,135.4, 132.5, 129.1, 129.0 (2C), 125.9, 123.3, 121.3 (2C), 119.2, 20.9.HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₆H₁₆ON₅ ⁺294.13494; Found 294.13495.HPLC (λ₂₅₄): 99.7%; t_(R): 8.456 min (method 4).

2-((4-amino-6-phenyl-1,3,5-triazin-2-yl)amino)phenol (MTF412)

Synthesized following the general procedure E using1-(benzo[d]oxazol-2-yl)guanidine (500 mg, 2.8 mmol), sodium hydride (60%suspension in oil, 123 mg, 3.1 mmol) and benzonitrile (0.29 mL, 2.8mmol) to afford the title compound as a light brown powder (53 mg, 7%).¹H NMR (400 MHz, Acetone-d6): δ 9.82 (br. s, 1H), 8.39 (d, J=7.1 Hz,2H), 8.31 (br. s, 1H), 7.83 (d, J=7.7 Hz, 1H), 7.58-7.52 (m, 1H),7.52-7.46 (m, 2H), 7.03-6.94 (m, 2H), 6.90-6.84 (m, 1H), 6.77 (br. s,2H). ¹³C NMR (101 MHz, Acetone-d6): δ 172.0, 168.5, 165.7, 148.9, 137.4,132.6, 129.1 (2C), 129.0 (2C), 128.5, 125.3, 123.1, 120.7, 118.3.HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₅H₁₄ON₅ ⁺280.11929; Found 280.11929.HPLC (λ₂₅₄): 98.9%; t_(R): 8.100 min (method 4).

2-((6-imino-4-phenyl-1,6-dihydro-1,3,5-triazin-2-yl)amino)-4-nitrophenol(MTF413)

Synthesized following the general procedure E using1-(6-nitrobenzo[d]oxazol-2-yl)guanidine (500 mg, 2.3 mmol), sodiumhydride (60% suspension in oil, 100 mg, 2.5 mmol) and benzonitrile (0.24mL, 2.3 mmol) to afford the title compound as a light yellow powder (62mg, 8%). ¹H NMR (400 MHz, DMSO-d6): δ 11.39 (br. s, 1H), 8.63 (d, J=9.0Hz, 1H), 8.36-8.30 (m, 2H), 8.26 (s, 1H), 7.80 (dd, J=9.0, 2.6 Hz, 1H),7.70 (d, J=2.6 Hz, 1H), 7.61-7.56 (m, 1H), 7.52 (dd, J=11.4, 4.4 Hz,2H), 7.45 (br. s, 2H). ¹³C NMR (101 MHz, DMSO-d6): δ 170.7, 167.2,164.2, 146.4, 141.5, 136.1, 134.5, 131.9, 128.5 (2C), 128.0 (2C), 118.9,115.4, 108.9. HRMS-ESI (m/z): [M+H]⁺ calc. for C₁₅H₁₃O₃N₆ ⁺325.10436;Found 325.10446. HPLC (λ₂₅₄): 99.1%; t_(R): 10.242 min (method 4).

2-((6-imino-4-(pyrazin-2-yl)-1,6-dihydro-1,3,5-triazin-2-yl)amino)phenol(MTF439)

Synthesized following the general procedure E using1-(benzo[d]oxazol-2-yl)guanidine (500 mg, 2.8 mmol), sodium hydride (60%oil suspension, 123 mg, 3.08 mmol) and pyrazinecarbonitrile (250 μL, 2.8mmol) to afford the title compound as a bright yellow powder (510 mg,65%). The biguanide was then dissolved in DMSO and stirred for 8 h atr.t. to give the desired product as a bright yellow powder. ¹H NMR (400MHz, DMSO-d6): δ 10.07 (br. s, 1H), 9.41 (s, 1H), 8.79 (s, 2H), 8.57 (s,1H), 7.88 (dd, J=8.0, 0.6 Hz, 1H), 7.53 (br. s, 1H), 7.41 (br. s, 1H),6.99-6.93 (m, 1H), 6.90 (dd, J=8.0, 1.4 Hz, 1H), 6.85-6.79 (m, 1H). ¹³CNMR (101 MHz, DMSO-d6): δ 168.7, 167.2, 164.6, 149.4, 148.4, 146.4,144.6, 144.5, 126.8, 124.4, 123.0, 119.2, 116.1. HRMS-ESI (m/z): [M+H]⁺calc. for C₁₃H₁₂ON₇ ⁺282.10978; Found 282.10977. HPLC (λ₂₅₄): 98.8%;t_(R): 4.209 min (method 4).

N—(N-(benzo[d]oxazol-2-yl)carbamimidoyl)pyrazine-2-carboximidamide(MTF440)

Synthesized following the general procedure E using1-(benzo[d]oxazol-2-yl)guanidine (500 mg, 2.8 mmol), sodium hydride (60%oil suspension, 123 mg, 3.08 mmol) and 3-pyridinecarbonitrile (292 mg,2.8 mmol) to afford the title compound as a bright yellow powder (510mg, 65%). ¹H NMR (400 MHz, DMSO-d6): δ 9.98 (s, 1H), 9.40 (d, J=1.4 Hz,1H), 8.73 (dd, J=4.8, 1.7 Hz, 1H), 8.54 (d, J=8.0 Hz, 1H), 8.34 (s, 1H),7.93 (dd, J=7.9, 1.4 Hz, 1H), 7.54 (ddd, J=8.0, 4.8, 0.5 Hz, 1H), 7.32(s, 2H), 6.98-6.93 (m, 1H), 6.91-6.88 (m, 1H), 6.86-6.80 (m, 1H). ¹³CNMR (101 MHz, DMSO-d6): δ 168.8, 167.0, 164.4, 152.1, 149.1, 148.1,135.2, 132.0, 126.8, 124.1, 123.6, 122.7, 119.1, 115.7. HRMS-ESI (m/z):[M+H]⁺ calc. for C₁₄H₁₃ON₆ ⁺281.11454; Found 281.11450. HPLC (2₂₅₄):99.7%; t_(R): 4.165 min (method 4).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(naphthalen-2-yl)-1,3,5-triazine-2,4-diamine)(MTF443)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2-naphthimidamide (345 mg, 1mmol) to afford the title compound as a white powder (299 mg, 87%). ¹HNMR (400 MHz, DMSO-d6): δ 9.30 (s, 1H), 8.87 (s, 1H), 8.36 (d, J=8.5 Hz,1H), 7.99 (dd, J=19.5, 10.4 Hz, 3H), 7.68 (d, J=7.8 Hz, 1H), 7.63-7.53(m, 2H), 7.48 (d, J=7.7 Hz, 1H), 7.29 (t, J=7.2 Hz, 1H), 7.25-7.11 (m,3H). ¹³C NMR (101 MHz, DMSO-d6): δ 170.19, 167.33, 165.53, 136.81,134.58, 134.17, 133.57, 132.40, 128.98, 128.66, 128.16, 127.74, 127.65(2C), 127.51, 126.55 (2C), 126.26, 124.75. HPLC (λ₂₈₀): Purity 97.3%;t_(R): 15.642 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(2,2-dimethyl-2H-chromen-6-yl)-1,3,5-triazine-2,4-diamine)(MTF444)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2,2-dimethyl-2H-chromene-6-carboximidamide(377 mg, 1 mmol) to afford the title compound as a white powder (241 mg,64%). ¹H NMR (400 MHz, DMSO-d6): δ 9.10 (s, 1H), 8.00 (d, J=8.5 Hz, 1H),7.95 (s, 1H), 7.61 (d, J=9.0 Hz, 1H), 7.41 (d, J=7.1 Hz, 1H), 7.27 (t,J=7.0 Hz, 1H), 7.16 (t, J=7.5 Hz, 1H), 6.99 (s, 2H), 6.81 (d, J=8.5 Hz,1H), 6.45 (d, J=9.9 Hz, 1H), 5.80 (d, J=9.8 Hz, 1H), 1.40 (s, 6H). ¹³CNMR (101 MHz, DMSO-d6): δ 169.67, 167.15, 165.33, 155.42, 136.86,133.39, 131.25, 129.26, 129.18, 128.51, 127.69, 126.41, 126.16 (2C),121.61, 120.26, 115.65, 76.97, 27.93 (2C). HPLC (λ₂₈₀): Purity 95.2%;t_(R): 15.767 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(3-nitrophenyl)-1,3,5-triazine-2,4-diamine)(MTF445)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3-nitrobenzimidamide (340 mg,1 mmol) to afford the title compound as a white powder (265 mg, 78%). ¹HNMR (400 MHz, DMSO-d6): δ 9.41 (s, 1H), 9.04 (s, 1H), 8.60 (d, J=6.8 Hz,1H), 8.39 (dd, J=8.2, 1.4 Hz, 1H), 7.79 (t, J=8.0 Hz, 1H), 7.64 (dd,J=7.9, 0.9 Hz, 1H), 7.39 (dd, J=7.8, 0.9 Hz, 1H), 7.36-7.17 (m, 4H). ¹³CNMR (101 MHz, DMSO-d6): δ 168.16, 167.23, 165.49, 147.97, 138.33,136.42, 133.75 (2C), 130.10, 128.35, 127.72, 126.82, 126.57, 125.93,122.27. HPLC (λ₂₈₀): Purity 95.3%; t_(R): 13.550 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(3,4,5-trimethoxyphenyl)-1,3,5-triazine-2,4-diamine)(MTF446)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3,4,5-trimethoxybenzimidamide(385 mg, 1 mmol) to afford the title compound as a white powder (358 mg,93%). ¹H NMR (400 MHz, DMSO-d6): δ 9.24 (s, 1H), 7.59 (s, 3H), 7.40 (d,J=7.7 Hz, 1H), 7.27 (t, J=7.5 Hz, 1H), 7.12 (dd, J=23.2, 15.8 Hz, 3H),3.79 (s, 6H), 3.73 (s, 3H). ¹³C NMR (101 MHz, DMSO-d6): δ 169.55,167.26, 165.29, 152.58 (2C), 140.36, 136.92, 133.81, 131.90, 128.71,127.66, 126.37, 126.11, 105.08 (2C), 60.11, 55.74 (2C). HPLC (λ₂₈₀):Purity 96.6%; t_(R): 10.367 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(benzo[d][1,3]dioxol-5-yl)-1,3,5-triazine-2,4-diamine)(MTF449)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)benzo[d][1,3]dioxole-5-carboximidamide(340 mg, 1 mmol) to afford the title compound as a white powder (247 mg,73%). ¹H NMR (400 MHz, DMSO-d6): δ 9.14 (s, 1H), 7.87 (d, J=8.1 Hz, 1H),7.68 (s, 1H), 7.61 (d, J=7.8 Hz, 1H), 7.39 (d, J=7.7 Hz, 1H), 7.27 (t,J=7.4 Hz, 1H), 7.17 (t, J=7.5 Hz, 1H), 7.08-6.97 (m, 3H), 6.10 (s, 2H).¹³C NMR (101 MHz, DMSO-d6): δ 169.41, 167.15, 165.34, 150.10, 147.37,136.77, 133.53, 130.76, 128.41, 127.66, 126.47, 126.21, 122.88, 107.97,107.44, 101.60. HPLC (λ₂₈₀): Purity 95.1%; t_(R): 10.767 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(2-chlorophenyl)-1,3,5-triazine-2,4-diamine)(MTF450)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2-chlorobenzimidamide (329 mg,1 mmol) to afford the title compound as a white powder (273 mg, 83%). ¹HNMR (400 MHz, DMSO-d6): δ 9.30 (s, 1H), 7.58 (dt, J=9.4, 4.8 Hz, 2H),7.51 (dd, J=7.8, 1.0 Hz, 1H), 7.45 (td, J=7.7, 2.0 Hz, 1H), 7.41 (dd,J=7.4, 1.3 Hz, 1H), 7.38 (d, J=6.8 Hz, 1H), 7.28-7.22 (m, 1H), 7.22-7.09(m, 3H). ¹³C NMR (101 MHz, DMSO-d6): δ 172.24, 166.83, 165.19, 137.48,136.34, 133.55, 130.93, 130.50, 130.47, 129.82, 128.16, 127.59, 127.08,126.87, 126.52. HPLC (λ₂₈₀): Purity 95.1%; t_(R): 10.517 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(p-tolyl)-1,3,5-triazine-2,4-diamine)(MTF451)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-4-methylbenzimidamide (309 mg,1 mmol) to afford the title compound as a white powder (179 mg, 58%). ¹HNMR (400 MHz, DMSO-d6): δ 9.15 (s, 1H), 8.14 (d, J=8.0 Hz, 2H), 7.61 (d,J=7.9 Hz, 1H), 7.41 (d, J=7.1 Hz, 1H), 7.26 (dd, J=8.8, 4.7 Hz, 3H),7.17 (t, J=7.5 Hz, 1H), 7.04 (s, 2H), 2.36 (s, 3H). ¹³C NMR (101 MHz,DMSO-d6): δ 170.14, 167.25, 165.46, 141.31, 136.80, 133.90, 133.44,128.86 (2C), 128.52, 127.86 (2C), 127.71, 126.51, 126.21, 21.09. HPLC(λ₂₈₀): Purity 96.7%; t_(R): 12.958 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(2-fluorophenyl)-1,3,5-triazine-2,4-diamine)(MTF452)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2-fluorobenzimidamide (313 mg,1 mmol) to afford the title compound as a white powder (191 mg, 61%). ¹HNMR (400 MHz, DMSO-d6): δ 9.26 (s, 1H), 7.87 (td, J=7.7, 1.6 Hz, 1H),7.59 (dd, J=7.9, 1.0 Hz, 1H), 7.56-7.49 (m, 1H), 7.39 (d, J=7.6 Hz, 1H),7.30-7.23 (m, 3H), 7.20-7.10 (m, 3H). ¹³C NMR (101 MHz, DMSO-d6): δ169.77 (d), 166.97, 165.31, 160.33 (d), 136.49, 133.54, 132.19 (d),131.13 (d), 128.26, 127.65, 126.92, 126.46, 125.89 (d), 124.09 (d),116.50 (d). HPLC (λ₂₈₀): Purity 95.4%; t_(R): 10.333 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(2,6-dichlorophenyl)-1,3,5-triazine-2,4-diamine)(MTF455)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2,6-dichlorobenzimidamide (364mg, 1 mmol) to afford the title compound as a white powder (320 mg,88%). ¹H NMR (400 MHz, DMSO-d6): δ 9.44 (s, 1H), 7.55 (dd, J=13.4, 7.9Hz, 3H), 7.45 (dd, J=9.0, 7.1 Hz, 1H), 7.33 (d, J=7.7 Hz, 1H), 7.30-7.22(m, 3H), 7.18 (t, J=7.1 Hz, 1H). ¹³C NMR (101 MHz, DMSO-d6): δ 170.75,166.93, 165.40, 136.69, 135.98, 133.90, 132.08 (2C), 130.63, 128.12(2C), 127.88, 127.50, 127.25, 126.83. HPLC (λ₂₈₀): Purity 95.8%; t_(R):10.150 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(3,5-dibromopyridin-4-yl)-1,3,5-triazine-2,4-diamine)(MTF456)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-3,5-dibromoisonicotinimidamide(454 mg, 1 mmol) to afford the title compound as a white powder (412 mg,91%). ¹H NMR (400 MHz, DMSO-d6): δ 9.56 (s, 1H), 8.84 (s, 2H), 7.57 (d,J=7.7 Hz, 1H), 7.39 (d, J=14.8 Hz, 2H), 7.31 (d, J=7.6 Hz, 1H), 7.25 (t,J=7.2 Hz, 1H), 7.20 (d, J=5.4 Hz, 1H). ¹³C NMR (101 MHz, DMSO-d6): δ170.93, 166.78, 165.27, 150.32 (2C), 146.43, 135.71, 133.98, 127.70,127.49, 127.35, 127.02, 119.05 (2C). HPLC (λ₂₈₀): Purity 95.1%; t_(R):9.700 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(6-bromobenzo[d][1,3]dioxol-5-yl)-1,3,5-triazine-2,4-diamine)(MTF458)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-6-bromobenzo[d][1,3]dioxole-5-carboximidamide(418 mg, 1 mmol) to afford the title compound as a white powder (300 mg,72%). ¹H NMR (400 MHz, DMSO-d6): δ 9.31 (s, 1H), 7.57 (d, J=7.6 Hz, 1H),7.38 (d, J=7.3 Hz, 1H), 7.28-7.23 (m, 2H), 7.22-7.11 (m, 4H), 6.12 (s,2H). ¹³C NMR (101 MHz, DMSO-d6): δ 172.43, 166.50, 165.08, 148.68,146.86, 136.29, 133.50, 132.49, 128.17, 127.63, 127.08, 126.58, 112.81,111.49, 109.98, 102.31. HPLC (λ₂₈₀): Purity 96.1%; t_(R): 10.475 min(method 12).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(3-aminophenyl)-1,3,5-triazine-2,4-diamine)(MTF460)

Synthesized following the general procedure B using3-amino-N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)benzimidamide (310 mg,1 mmol) to afford the title compound as a yellow powder (241 mg, 78%).¹H NMR (400 MHz, DMSO-d6): δ 9.06 (s, 1H), 7.61 (dd, J=7.9, 1.2 Hz, 1H),7.49 (s, 1H), 7.43 (d, J=7.8 Hz, 2H), 7.27 (td, J=7.7, 1.3 Hz, 1H), 7.17(t, J=7.6 Hz, 1H), 7.09 (t, J=7.8 Hz, 1H), 6.98 (s, 2H), 6.71 (dd,J=7.9, 1.4 Hz, 1H), 5.18 (s, 2H). ¹³C NMR (101 MHz, DMSO-d6): δ 170.95,167.23, 165.44, 148.52, 137.33, 136.87, 133.20, 128.59, 128.55, 127.72,126.43, 126.12, 116.94, 115.86, 113.34. HPLC (λ₂₈₀): Purity 97.8%;t_(R): 7.167 min (method 2).

1,1′-((((disulfanediylbis(2,1-phenylene))bis(azanediyl))bis(6-amino-1,3,5-triazine-4,2-diyl))bis(3,1-phenylene))bis(ethan-1-one)(MTF462)

Synthesized following the general procedure B using3-acetyl-N—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)benzimidamide (337 mg,1 mmol) to afford the title compound as a white powder (225 mg, 67%). ¹HNMR (400 MHz, DMSO-d6): δ 9.32 (s, 1H), 8.83 (s, 1H), 8.46 (d, J=7.4 Hz,1H), 8.13 (d, J=7.8 Hz, 1H), 7.64 (t, J=7.1 Hz, 2H), 7.40 (d, J=7.1 Hz,1H), 7.23 (dt, J=15.3, 8.0 Hz, 4H), 2.61 (s, 3H). ¹³C NMR (101 MHz,DMSO-d6): δ 197.57, 169.41, 167.28, 165.49, 137.06, 136.86, 136.62,133.74, 132.14, 131.24, 128.80, 128.46, 127.69, 127.32, 126.67, 126.40,26.77. HPLC (λ₂₈₀): Purity 96.9%; t_(R): 10.908 min (method 2).

N2,N2′-(disulfanediylbis(2,1-phenylene))bis(6-(2-nitrophenyl)-1,3,5-triazine-2,4-diamine)(MTF463)

Synthesized following the general procedure B usingN—(N-(benzo[d]thiazol-2-yl)carbamimidoyl)-2-nitrobenzimidamide (340 mg,1 mmol) to afford the title compound as a white powder (278 mg, 82%). ¹HNMR (400 MHz, DMSO-d6): δ 9.29 (s, 1H), 7.92 (d, J=7.6 Hz, 1H), 7.86 (d,J=7.3 Hz, 1H), 7.77 (t, J=7.3 Hz, 1H), 7.70 (t, J=7.6 Hz, 1H), 7.57 (d,J=7.8 Hz, 1H), 7.32 (d, J=7.6 Hz, 1H), 7.28-7.07 (m, 4H). ¹³C NMR (101MHz, DMSO-d6): δ 170.32, 166.77, 165.14, 149.12, 136.11, 133.49, 132.58,132.34, 130.89, 130.51, 128.06, 127.58, 127.11, 126.63, 123.88. HPLC(λ₂₈₀): Purity 98.9%; t_(R): 10.350 min (method 2).

Cell Cultures

Fresh sterile tissues were obtained from surgical waste from patientsdiagnosed with metastatic melanoma at the Nice CHU hospital. Epidermalcell suspensions were obtained from foreskins of Caucasian children byovernight digestion in phosphate-buffered saline containing 0.5% dispasegrade II at 4° C., followed by a 20 min digestion with 0.05%trypsin-0.02% EDTA in phosphate-buffered saline (V/V) at 37° C. Humanprimary melanocytes were grown in MCDB153 medium supplemented with 2%FCS, 0.4 μg/ml hydrocortisone, 5 μg/ml insulin, 16 nM phorbol-12myristate 13-acetate, 1 ng/ml basic fibroblast growth factor, 10 μg/mlbovine pituitary extract and penicillin/streptomycin (100 U/ml/50μg/ml). Human primary keratinocytes were cultured in KSFM medium inwhich 0.1 ng/ml epidermal growth factor, 15 μg/ml bovine pituitaryextract and penicillin/streptomycin (100 U/ml/50 μg/ml) were added.Human primary fibroblasts and melanoma cells, derived from thecorresponding dermis, were grown in DMEM 7% Foetal Calf Serum (FCS) andpenicillin/streptomycin (100 U/ml/50 μg/ml). Written informed consentwas obtained from each patient included in this study, and the study wasapproved by the hospital ethics committee (Nice Hospital Center andUniversity of Nice Sophia Antipolis, no. 210-2998).

Different melanoma cell lines were purchased from the American TissueCulture Collection: A375 and WM9 cells are mutated on B-Raf, CDKN2A andPTEN proteins; SKmel28 cells are mutated on B-Raf and P53 proteins;1205Lu cells are mutated on B-Raf and PTEN proteins; G-361 cells aremutated on B-Raf and CDKN2A proteins; C8161 cells are mutated on B-Rafand K-Ras proteins; WM3912 cells are mutated on B-Raf and CDKN2Aproteins; MeWo cells are mutated on P53 and CDKN2A proteins; WM3918presents no mutations on characteristic proteins.

Resistant melanoma cell lines A375 and SKMel28 were a gift fromProfessor P. Marchetti and described in Corazao-Rozas et al. (2013).Cells were grown in RPMI 1640 or in DMEM supplemented with 10% of FoetalCalf Serum (FCS) and penicillin/streptomycin (100 U/ml/50 mg/ml) at 37°C. and 5% CO2.

Trypan Blue Exclusion Assay

For trypan blue staining, 200 mL of cells (melanoma and normal humancells) were aseptically transferred to a 1.5 mL clear Eppendorf tube andincubated for 3 min at room temperature with an equal volume of 0.4%trypan blue solution. Viable cells were counted and the results areexpressed as the percentage of the value of control cells. All theexperiments are performed 3 times in triplicate.

Viability Test

A375 Sensitive cells were treated with different concentrations of thesynthesized molecules (5.0 μM or 10 μM) for different times (24 hr or 48hr) or with DMSO (Control) for 12 or 48 hr. At the end of thestimulation, viable cells were counted using the trypan blue dyeexclusion method. The results are normalized as percentages compare tothe control. The results are shown in Table 1. They show that compoundsof formula (I), (II) and (III) induce a decrease of the viability ofmelanoma cells.

Kinetic of CRO15

A375 Sensitive cells were treated with 5 μM of CRO15 for different times(2 hr, 4 hr, 6 hr, 8 hr, 12 hr, 24 hr or 48 hr) or with 5 μM of PLX4032(B-Raf inhibitor) for 48 hr or with DMSO (Control) for 48 hr. At the endof stimulation, viable cells were counted using the trypan blue dyeexclusion method. The results are normalized as percentages compare tothe control and data means±SEM of three independent experimentsperformed in triplicate *p<0.05; **p<0.01; ***p<0.001. The results areshown in FIG. 1A. They show that CRO15 induces a decrease of theviability of melanoma cells and that the decrease of the viability after48 hr is more important with CRO15 than with PLX4032.

IC50 of CRO15

A375 Sensitive cells were treated with different concentrations of CRO15(0.5 μM, 2.5 μM, 5.0 μM, 7.5 μM, 50 μM) for 48 hr or with 5 μM ofPLX4032 (B-Raf inhibitor) for 48 hr or with DMSO (Control) for 48 hr. Atthe end of the stimulation, viable cells were counted using the trypanblue dye exclusion method. The results are normalized as percentagescompare to the control and data means±SEM of three independentexperiments performed in triplicate *p<0.05; **p<0.01; ***p<0.001. IC50of CRO15 have been determined at 3.75 μM at 48 hr. The results are shownin FIG. 1B. They show that there is a dose-response relationship forCRO15 on melanoma cells viability.

Viability Test with CRO15

Different melanoma cell lines with various mutations, patient cells withvarious mutations and normal human cells, were treated with 5 μM ofCRO15 for 48 hours or DMSO (Control). At the end of stimulation, viablecells were counted using the trypan blue dye exclusion method. Theresults are normalized as percentages compare to the control and datameans±SEM of three independent experiments performed in triplicate*p<0.05; **p<0.01; ***p<0.001. The results are shown in FIG. 1C(melanoma cell lines with various mutations) and 1D (patient cells withvarious mutations) and 1E (normal human cells). They show that CRO15induces a decrease of the viability of different melanoma cell lineswith various mutations and different patient cells with variousmutations, while CRO15 is not toxic for normal cells.

Viability Test of CRO15 on Resistant Melanoma Cells

A375 sensitive and resistant melanoma cells were treated with 5 μM ofCRO15 for 48 hr or with 5 μM of PLX4032 for 48 hr or with DMSO(Control). At 48 hours, viable cells were counted using the trypan bluedye exclusion method. The results are normalized as percentages compareto the control and data means±SEM of three independent experimentsperformed in triplicate *p<0.05; **p<0.01; ***p<0.001. The results areshown in FIG. 4A. They show that CRO15 induces a decrease of the cellviability in both sensitive and resistant melanoma cells and that thedecrease of the viability is more important with CRO15 than withPLX4032.

Viability Test of CRO15 on Double Resistant Cells to BRAF and MEKInhibitors

Melanoma cell line DR6 resistant cells to B-Raf inhibitor(Vemurafenib=PLX4032) and to MEK inhibitor (Cobimetinib) were treatedwith DMSO (control), with 5 μM or 10 μM of CRO15 or with a combinationof 1 μM of vemurafenib and 0.5 μM of cobimetinib. After 24 hours ofstimulation, viable cells were counted using the trypan blue dyeexclusion method. The results are normalized as percentages compare tothe control. Error bars represents ±SEM of triplicate.

The results are shown in FIG. 5. They show that CRO15 induces a decreaseof the cell viability of melanoma cell line resistant to B-Raf inhibitor(PLX4032) and to MEK inhibitor (Cobimetinib).

Western Blot Assays

Western blot analyses were performed as described (Lehraiki et al.,2014). Proteins were extracted in buffer containing 50 mmol/l Tris-HCl(pH 7.5), 15 mmol/1, NaCl, 1% Triton X-100, and 1× protease andphosphatase inhibitors. Briefly, cell lysates (30 mg) were separated bySDS-PAGE, transferred onto a polyvinylidene fluoride membrane(Millipore), and then exposed to the appropriate antibodies. Proteinswere visualized with the ECL System from Amersham. The western blotanalyses shown are representative of at least three independentexperiments.

Western Blot analysis: A375 Sensitive cells were treated with 5 μM ofCRO15 for different time (6 hr, 12 hr or 24 hr) or with DMSO (Control)for 24 hr.

The results are shown in FIG. 2. They show that CRO15 induces anactivation of AMPK.

In Vivo Murine Cancer Model

Animal experiments were carried out in accordance with the Declarationof Helsinki and were approved by a local ethical committee CIEPAL(Comité Institutionnel d'Ethique Pour l'Animal de Laboratoire-Azur).Female immune-deficient BALB/c nu/nu (nude) mice were obtained at 5weeks of age from Envigo Laboratory (Gannat, France). Nude mice wereinoculated subcutaneously with A375 sensitive or resistant melanomacells (1.0×106 cells/mouse). After tumor apparition (±5 days), animalsreceived intraperitoneal injection of Labrafil (Control), PLX4032 (0.7mg/mouse/day) or CRO15 (0.7 mg/mouse/day) dissolved in Labrafil. Thegrowth tumor curves were determined by measuring the tumor volume usingthe equation V=(L*W2)/2 (V=tumor volume, W=tumor width, L=tumor length).At the end of the experiment, mice were euthanized by cervicaldislocation and tumors were taken for western blot andimmunofluorescence experiments (LC3, Cleaved Caspase 3). TUNEL assay wasperformed using the In Situ Cell Death Detection Kit (Roche, Meylan,France). The results are shown in FIGS. 3A and 3B (A375 sensitivemelanoma cells), and 4B and 4C (A375 resistant melanoma cells). Theyshow that CRO15 reduces both the tumor volume and weight of miceinoculated with both sensitive and resistant melanoma cells.

Xenograft—A375 Resistant cells (Riv)

In Vivo Murine Cancer Model Animal experiments were carried out inaccordance with the Declaration of Helsinki and were approved by a localethical committee CIEPAL (Comite Institutionnel d'Ethique Pour l'Animalde Laboratoire-Azur). Female immune-deficient BALB/c nu/nu (nude) micewere obtained at 5 weeks of age from Envigo Laboratory (Gannat, France).Nude mice were inoculated subcutaneously with A375 resistant melanomacells (1.0×106 cells/mouse). After tumor apparition (+5 days), animalsreceived intraperitoneal injection of Labrafil (Control), PLX4032 (0.7mg/mouse/day) or MTF319 (0.7 mg/mouse/day) dissolved in Labrafil. Thegrowth tumor curves were determined by measuring the tumor volume usingthe equation V=(L*W2)/2 (V=tumor volume, W=tumor width, L=tumor length).At the end of the experiment, mice were euthanized by cervicaldislocation.

The results are shown in FIGS. 6A and 6B. They show a decrease in tumorvolume when mice were injected with MTF319. The tumor weight alsoindicates that MTF319 had an effect on the tumor growth.

CRO15 Inhibits Tumor Growth of Murine Melanoma BP Cells Allografted intoC57BL6 Mice

In Vivo Murine Cancer Model Animal experiments were carried out inaccordance with the Declaration of Helsinki and were approved by a localethical committee CIEPAL (Comite Institutionnel d'Ethique Pour l'Animalde Laboratoire-Azur). Female C57BL6/J mice were obtained at 5 weeks ofage from Envigo Laboratory (Gannat, France). Mice were inoculatedsubcutaneously with BP melanoma cells (1×10⁶ cells/mouse). After tumorapparition (+5 days), animals received intraperitoneal injection ofLabrafil (vehicle) or CRO15 (0.7 mg/mouse/day). The growth tumor curveswere determined by measuring the tumor volume using the equationV=(L*W2)/2, (V=tumor volume, W=tumor width, L=tumor length). At the endof the experiment, mice were euthanized by cervical dislocation.

The results are shown in FIG. 7A. The bars indicate the mean±SEM.*p<0.05; **p<0.01. They show that immunocompetent mice (C57BL/6)subcutaneously injected with murine melanoma cells (BP cells) showed notumor growth when treated daily with CRO15 compared with those treatedwith vehicle.

To study weither melanoma cells WM9 would develop resistance to CRO15,these cells were kept in culture during 8 weeks in presence ofincreasing concentrations of either, DMSO (control), PLX4032, CRO15 orMTF255. The starting concentration was 0.2 μM. Every 2 passages, drugconcentration was slightly increased (+0.2 μM) until reaching resistanceto PLX4032 when channeling with 10 μM as observed in the FIG. 7B.

Along with non-treated WM9S (naive cells), drug-treated WM9S were thenstimulated with 10 μM of each drug during 48 hrs. Viable cells werecounted using the trypan blue dye exclusion method. The results arenormalized as percentages compare to the control (DMSO). Error barsrepresents ±SEM of triplicate.

The results are shown in FIG. 7B. They show that that PLX-treated WM9became resistant to PLX4032 while CRO15 is still able to induce celldeath in CRO15-treated WM9.

TABLE 1 Viab Viab Viab Viab A375 A375 A375 A375 10 μM/ 5 μM/ 10 μM/ 5μM/ ID Structure 24 h 24 h 48 h 48 h CRO15

23% 36% 2.6% 27% MTF 232

33.9% ±2 83.3% ±5.3 16.4 ±4.9% 24.9% ±8.5 MTF 233

7% ±0.6 13.9% ±3.6 4.7% ±1.7 8.9% ±2.1 MTF 234

9.6% ±3.2 29.6% ±6.1 7.8% ±3.6 17.4% ±3.6 MTF 242

13.4% ±4.5 29.2% ±0.0 2.9% ±4 8.1% ±3.1 MTF 243

12.1% ±3.1 22.1% ±3.8 2.7% ±1.7 7% ±11.1 MTF 244

84.9% ±9.9 81.6% ±22.6 68.4% ±13.4 68% ±26 MTF 245

83% ±16.3 95% ±9.2 79.6% ±7.8 100% ±8.5 MTF 246

94% ±58 137% ±33.7 115% ±26 91% ±7.8 MTF 247

122% ±79 100% ±33 100% ±49 135% ±39 MTF 248

80% ±7.2 100% ±31 138% ±59 100% ±16 MTF 249

14.4% ±1 25.4% ±8.1 8.9% ±2.9 10.6% ±6.7 MTF 250

12% ±1.5 25.4% ±4.2 2.4% ±1.2 8.9% ±5.6 MTF 251

45.1% ±3.1 48.9% ±3.6 19.5% ±7.8 44.7% ±2.9 MTF 252

12.9% ±1 8.6% ±3 2.6% ±1 5.2% ±4.6 MTF 253

14.9% ±2.5 27.8% ±7.6 4.1% ±3.5 6.9% ±5 MTF 254

100% ±31 100% ±23 100% ±12 100% ±55 MTF 255

3.2% ±1.7 IC50 = 2.3 μM 26.1% ±7.1 3.8% ±0.6 4.7% ±5.3 MTF 256

66% ±3.2 93.9% ±2.3 95.7% ±20 60% ±7.1 MTF 257

106% ±18 100% ±10 62% ±11 80.5% ±7.1 MTF 259

98% ±24 157% ±1.7 96.5% ±11 95.7% ±26.3 MTF 260

76.7% ±21 57% ±2.5 78% ±65 32% ±24 MTF 261

71.9% ±6.4 24.6% ±11 115% ±1.4 63.6% ±9.9 MTF 262

40.8% ±9.1 61.4% ±15.7 27.3% ±5.3 43.7% ±17.6 MTF 263

100% ±7.6 89.5% ±8.4 66% ±8 91% ±6.8 MTF 264

89% ±19 87% ±15 99% ±28 94.9% v46 MTF 265

84% ±3.8 91% ±8.7 60% ±8.4 100% ±10 MTF 267

134% ±5.4 113% ±11.9 73.3% ±39 113% ±29 MTF 268

146% ±13.7 67% ±6.1 122% ±43 47% ±5.5 MTF 272

88% ±10 100% ±6.2 54.6% ±32 81% ±21 MTF 273

100% ±6.6 46% ±2.9 100% ±12 48% ±8.2 MTF 274

84% ±11 83% ±5.9 89% ±20 93% ±15 MTF 276

73.9% ±24.4 91.6% ±5.1 64.9% ±7.2 95.3% ±13.9 MTF 277

76.4% ±5.2 77.6% ±16 85.8% ±10 88% ±12 MTF 281

57.4% ±4.9 79% ±15 78% ±15 112% ±2 MTF 283

100% ±9 100% ±10 150% ±4 56% ±5.6 MTF 284

104.1% ±11.4 80.1% ±3.1 76.9% ±9.2 117.3% ±13.1 MTF 285

77.8% ±8 45% ±4.5 78.7% ±3.8 100% ±8 MTF 286

65.3% ±7.3 48% ±2.3 120% ±11.5 155% ±16 MTF 287

53.7% ±3.8 60.9% ±11.7 69.5% ±14.2 86.7% ±19.8 MTF 288

70% ±4.6 50% ±10 53.5% ±14.9 96.7% ±17 MTF 289

78.7% ±29 81.1% ±7.2 58.2% ±12.5 90.4% ±26.5 MTF 290

50.5% ±5.6 43.4% ±8.3 14.4% ±2.4 21.6% ±15.9 MTF 291

100% ±11 100% ±12 41.3% ±1.8 95.2% ±15.9 MTF 292

141.9% ±12.9 88.1% ±8.9 71.9% ±8 83.3% ±18.7 MTF 295

66.1% ±7.2 108.9% ±18.9 39.3% ±4.2 37.3% ±15 MTF 296

21.1% ±9.5 39.4% ±7.1. 6.8% ±6.1 11.7% ±6.8 MTF 297

97.9% ±8.1 86.2% ±7.5 24.9% ±7.2 59.8% ±5.5 MTF 298

87.5% ±5.1 97.2% ±13 54.7% ±21.8 73.6% ±15.1 MTF 299

72.8% ±14.6 91.1% ±7.2 46.2% ±2.8 76.6% ±32.5 MTF 300

50.8% ±6.4 96% ±9.3 35.9% ±6 60% ±16.1 MTF 301

106.4% ±14.2 74% ±15 31.6% ±20 47% ±23 MTF 302

94.8% ±3.5 104.6% ±9 39.3% ±8.7 52.3% ±24.3 MTF 303

66.1% ±6.1 85.6% ±23.2 39.1% ±16 44.2% ±4.9 MTF 305

5.5% ±0 11% ±4.6 4.7% ±2.5 20.7% ±13.1 MTF 316

103.8% ±13.3 124.5% ±9.3 48.2% ±8.5 69.6% ±37.5 MTF 317

108.6% ±6.6 106.4% ±9.5 70% ±17.7 58.7% ±21.1 MTF 318

20.5% ±0.6 74.1% ±3.8 6.3% ±3.2 21.6% ±5 MTF 319

8.6% ±4 8.6% ±2.1 0.6% ±0.6 3.1% ±1.2 MTF 320

10.8% ±2.5 18.3% ±3.1 8.5% ±1.5 11.5% ±1.5 MTF 321

53.8% ±5.8 59.7% ±2.1 32.1% ±9.1 34.5% ±1.4 MTF 322

26.9% ±2.6 57.4% ±4 5.7% ±2 12.1% ±1.5 MTF 323

47.4% ±1.5 34.6% ±4.6 8.3% ±3.1 23.5% ±3.2 MTF 324

12.9% ±3 44.1% ±7.5 8.2% ±0.7 13.3% ±3.25 MTF 325

71% ±1 58.1% ±0 29.7% ±3.2 30.9% ±5.7 MTF 326

12.9% ±0 27.4% ±0.7 6.7% ±3.1 27.9% ±0.7 MTF 327

23.7% ±0.6 19.4% ±1.7 14.5% ±4.6 14.5% ±1 MTF 328

27.8% ±6.4 50% ±8.2 3.4% ±3 9.4% ±4.4 MTF 329

74.2% ±6.1 96.2% ±3.5 22.5% ±13.1 69.7% ±2.1 MTF 330

60.1% ±10.7 122.7% ±17 41.6% ±9.1 73.9% ±3.5 MTF 331

150.8% ±3.5 104.5% ±2.8 77.4% ±16.3 82.9% ±4.2 MTF 332

104.5% ±8.2 122% ±9.2 65.4% ±6.4 87.6% ±6.4 MTF 333

28.7% ±3.2 94.9% ±0.9 24.1% ±3.9 35.4% ±1 MTF 342

32.2% ±5.7 48.6% ±5.5 8.6% ±3.2 8.6% ±3.2 MTF 343

34.8% ±5.5 67.7% ±21 24.1% ±14.2 52.9% ±13.4 MTF 344

38.7% ±7.8 84.3% ±0.7 26.1% ±11.6 26.1% ±11.6 MTF 345

17.1% ±2.8 25.9% ±2.5 1% ±0.6 1.8% ±0.6 MTF 346

43.2% ±0 66.9% ±1.4 12.9% ±7.2 10.6% ±5.6 MTF 347

27.6% ±2.1 26.9% ±2.1 6.8% ±3.5 6.6% ±3.2 MTF 348

110% ±9.2 101.8% ±11 35.8% ±4.9 79.6% ±44.5 MTF 379

95.7% ±14.8 148.2% ±10.3 86.2% ±0 76.9% ±12.7 MTF 380

36.2% ±6 47.3% ±5.7 2.3% ±1 4.9% ±1.2 MTF 381

90.6% ±13.2 88.3% ±8.5 61.4% ±24.4 61.6% ±31.8 MTF 382

17.4% ±1.5 14.7% ±5.5 4.5% ±3.1 1.8% ±0.7 MTF 383

41.6% ±6.4 43.1% ±11.5 7.1% ±6.4 40.6% ±23.3 MTF 384

93.3% ±4.7 76.1% ±7.5 72.2% ±10.6 119.3% ±13.4 MTF 385

86.6% ±11.5 52.3% ±6.4 81.5% ±10.6 68.1% ±3.5 MTF 386

18.1% ±5.3 28.9% ±8.7 3.7% ±0.6 6.6% ±3.5 MTF 387

67.8% ±1.5 70.1% ±14.4 55.6% ±14 56.2% ±5 MTF 388

55.7% ±7.5 61.4% ±15.5 18.4% ±2.1 65.1% ±7.8 MTF 389

75.8% ±4 74.6% ±2.6 76.4% ±11.3 85.6% ±13.4 MTF 394

4.5% ±1 13.1% ±1.2 1.1% ±0.6 4.4% ±1.2 MTF 396

7.5% ±1.2 19.2% ±5.1 0% ±0 12.1% ±4.9 MTF 397

13.4% ±1 35.4% ±3.1 7.3% ±2.1 33.7% ±4 MTF 398

91% ±9.5 91.9% ±2.3 108.6% ±11.3 132.4% ±0.7 MTF 373

38.6% ±5.7 34.3% ±2.1 22.8% ±3.2 22.5% ±10.6 MTF 374

24.0% ±7.1 47.0% ±7.3 27.7% ±9.2 69.2% ±0.0 MTF 375

30.9% ±8.7 47.2% ±2.1 15.5% ±0.7 29.5% ±3.5 MTF 376

24.3% ±7.6 44.7% ±5.7 20.9% ±7.5 38.9% ±2.1 MTF 377

22.7% ±9.2 27.3% ±3.5 33.5% ±4.2 34.4% ±2.1 MTF 443

13.8% ±6.4 42.3% ±7.1 7.8% ±4.2 10.2% ±4.0 MTF 445

72.4% ±7.4 62.6% ±10.3 16.3% ±0.0 35.2% ±10.6 MTF 446

16.3% ±1.5 62.6% ±3.5 1.8% ±1.4 13.0% ±6.8 MTF 449

59.1% ±8.5 74.2% ±14.6 53.0% ±5.0 67.8% ±23.3 MTF 450

38.3% ±15.0 38.6% ±17.1 49.9% ±17.6 54.8% ±16.4 MTF 451

10.6% ±4.9 26.1% ±7.5 12.5% ±9.3 23.9% ±17.7 MTF 452

22.3% ±11.6 43.6% ±1.5 40.3% ±6.8 60.5% ±14.0 MTF 455

32.3% ±0.6 89.0% ±13.1 69.5% ±17.1 108.1% ±9.9 MTF 456

52.3% ±3.6 66.5% ±1.5 76.1% ±30.8 87.5% ±23.4 MTF 458

46.5% ±9.9 58.7% ±3.2 103.7% ±7.2 86.8% ±11.5 MTF 460

64.5% ±12.1 61.9% ±6.1 38.6% ±2.8 69.5% ±7.1 MTF 462

4.8% ±17.4 101.3% ±37.3 44.1% ±6.9 80.0% ±17.7 MTF 463

56.1% ±5.2 80.0% ±6.8 43.4% ±7.1 74.3% ±21.4

The invention claimed is:
 1. A compound of formula (II) or (III)

wherein Y′ is —SR₄ or —OR₅, R₄ is selected from C₁-C₆ alkyl andprotecting groups selected from any group linked by a disulfidefunction, thioesters, alkyl, alkenyl and alkynyl thioethers, benzylthioethers, alkylarylmethyl thioethers, and triarylmethylthioethers; R₅is selected from protecting groups selected from esters, silylatedethers, alkoxymethyl ethers, benzyl ethers tetrahydropyranyl ethers,pentoses, and hexoses; each R′ is independently selected from halogen,C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, —OH, —NR″R′″, —NO₂, —CN and—(CO)—R; n is 0 to 4; R₃ is selected from C₁-C₆ alkyl, C₃-C₆ cycloalkyl,C₁-C₆ haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, heterocyclyl having 5 to10 ring atoms, aryl having 6 to 10 ring atoms, heteroaryl having 5 to 10ring atoms and C₇-C₁₆ aralkyl, said alkyl, cycloalkyl, haloalkyl,alkenyl, alkynyl, heterocyclyl, aryl, heteroaryl and aralkyl beingoptionally substituted with one or more substituents independentlyselected from halogen, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆haloalkyl, —OH,—NR″R″, —NO₂, —CN and —(CO)—R; each R is independently selected from H,C₁-C₆ alkyl, C₁-C₆ alkoxy and —NR″R″ ‘; each R″ and R’ is independentlyselected from H and C₁-C₆ alkyl.
 2. The compound of claim 1, wherein Y′is —SR₄, R₄ being selected from C₁-C₆ alkyl; each R′ is independentlyselected from halogen and C₁-C₆ alkoxy; n is 0 to 1; R₃ is selected fromC₁-C₆ haloalkyl, aryl having 6 to 10 ring atoms, heteroaryl having 5 to10 ring atoms, and C₇-C₁₆ aralkyl, said aryl, heteroaryl and aralkylbeing optionally substituted with one or more substituents independentlyselected from halogen, C₁-C₆ alkoxy, and —CN.
 3. The compound of claim1, wherein the compound is


4. The compound of claim 1, wherein the compound is selected from


5. A pharmaceutical composition comprising a compound of claim 1, and apharmaceutically acceptable carrier.